Tissue-derived matrikine compositions and methods therefor

ABSTRACT

A composition for topical administration to an epithelium is disclosed. The composition comprises a deconstructed matrisome including one or more enzymatically fragmented peptides derived from biological tissue. The composition further comprises one or more pharmaceutically acceptable or cosmetically acceptable excipients. The deconstructed matrisome may be fragmented through degradation by one or more enzymes to produce the one or more peptides. The one or more peptides may be configured to retain cell signaling ability, thereby promoting one or more of tissue homeostasis, tissue repair, and tissue regeneration. The composition may be used to treat a tissue such as skin tissue exhibiting scarring, acne, eczema, psoriasis, and other skin conditions.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of priority to U.S. ProvisionalApplication No. 63/075,638 entitled “Tissue-Derived MatrikineCompositions and Methods Therefor,” filed Sep. 8, 2020, which isincorporated herein by reference in its entirety.

TECHNICAL FIELD

This present disclosure relates generally to compositions comprisingmatrisome components and related methods. The disclosed compositions andmethods may be utilized, for example, to treat scars and other skinconditions, such as acne, eczema, and psoriasis.

BACKGROUND

Dermal tissue damage affects 500+ million people each year worldwide,while minimal options for effective topical treatment are commerciallyavailable. Current top-selling over-the-counter scar treatment productsare limited to plant-based or chemical ingredients with no active repairor regenerative signaling capacity. There is a lack of products withbioactive ingredients that offer proven support for skin repair—asignificant market gap waiting to be filled.

Existing products to treat scar repair incorporate herbal or foodextracts or silicone-based or other synthetic ingredients but showlimited data around proven regenerative bioactivity or the ability totailor to specific skin condition, ethnicities, ages, etc. to facilitate“personalized” product offerings. There is a significant marketopportunity for a topical skin bioactive that can prevent andeffectively reduce redness and scars after injury by replicating naturalregenerative signals.

In its native environment, extracellular matrix (ECM) is a scaffold withtissue-specific cues (e.g., molecular, structural, biomechanical) thatprovides structure for cell maintenance and growth and mediates cellproliferation, differentiation, gene expression, migration, orientation,and assembly. ECM comprises an interlocking mesh of components includingbut not limited to viscous proteoglycans (e.g., heparin sulfate, keratinsulfate, and chondroitin sulfate) that provide cushioning, collagen andelastin fibers that provide strength and resilience, and solublemultiadhesive proteins (e.g., fibronectin and laminin) that bind theproteoglycans and collagen fibers to cell receptors. Nativeextracellular matrix also commonly includes hyaluronic acid and cellularadhesion molecules (CAMs) such as integrins, cadherins, selectins, andimmunoglobulins.

The complexity of the ECM has proven difficult to recapitulate in itsentirety outside of its native environment. Mimicking ECM structureusing synthetic biomaterials or mimicking composition by adding purifiedECM components is possible. While offering structural mimics, syntheticbiomaterials can alter cell behavior (i.e., proliferation,differentiation, gene expression, migration, orientation, and assembly)in vitro and potentially generate cytotoxic by-products at the site ofimplantation, leading to poor wound healing or an inflammatoryenvironment.

Further, the ECM of each type of tissue may comprise a different mixtureof components, concentrations of components, and/or properties that aresuited to the tissue's unique set of roles and may signal unique cellactivity. Further, disease states in tissues may be associated withspecific alterations in the biochemical composition, structure, andbiomechanics of the ECM environments.

As such, it would be advantageous to have a topical composition capableof activating regenerative bioactivity in tissue-specific and/orpopulation-specific manners by recapitulating in vivo nicheenvironments.

SUMMARY

This summary is provided to comply with 37 C.F.R. § 1.73. It issubmitted with the understanding that it will not be used to interpretor limit the scope or meaning of the present disclosure.

Embodiments of the invention are directed to a composition for topicaladministration to an epithelium, the composition comprising: adeconstructed matrisome including one or more enzymatically fragmentedpeptides derived from a at least one biological tissue; and one or morepharmaceutically acceptable or cosmetically acceptable excipients,wherein the deconstructed matrisome is in an amount from about 0.1% byweight to about 15% by weight of the composition; and wherein the one ormore enzymatically fragmented peptides are configured to retain cellsignaling ability, thereby promoting one or more of tissue homeostasis,tissue repair, and tissue regeneration.

Embodiments of the invention are directed to a method of promotingrepair or regeneration in a target tissue, the method comprising:topically administering a composition to an epithelium of the targettissue, the composition comprising: a deconstructed matrisome includingone or more enzymatically fragmented peptides derived from at least onebiological tissue, and one or more pharmaceutically acceptable orcosmetically acceptable excipients, wherein the deconstructed matrisomeis in an amount from about 0.1% by weight to about 15% by weight of thecomposition; and wherein the one or more enzymatically fragmentedpeptides are configured to retain cell signaling ability, therebypromoting one or more of tissue homeostasis, tissue repair, and tissueregeneration in the target tissue.

Embodiments of the invention are directed to a method of increasingkeratin gene expression in a target tissue, the method comprising:topically administering a composition to an epithelium of the targettissue, the composition comprising: a deconstructed matrisome includingone or more enzymatically fragmented peptides derived from at least onebiological tissue, and one or more pharmaceutically acceptable orcosmetically acceptable excipients, wherein the deconstructed matrisomeis in an amount of about 0.1% by weight to about 15% by weight of thecomposition; and wherein the one or more enzymatically fragmentedpeptides are configured to retain cell signaling ability, therebyreducing or preventing one or more of laxity, wrinkling, and sagging inthe target tissue.

Embodiments of the invention are directed to a method of reducing dermalredness in a target tissue, the method comprising: topicallyadministering a composition to an epithelium of the target tissue, thecomposition comprising: a deconstructed matrisome including one or moreenzymatically fragmented peptides derived from at least one biologicaltissue, and one or more pharmaceutically acceptable or cosmeticallyacceptable excipients, wherein the deconstructed matrisome is in anamount of about 0.1% by weight to about 15% by weight of thecomposition; and wherein the one or more enzymatically fragmentedpeptides are configured to retain cell signaling ability, therebypromoting healing or recovery in the target tissue from one or more ofscar formation, a wound, and a burn.

Embodiments of the invention are directed to a method of lowering the pHof a surface of a target tissue, the method comprising: topicallyadministering a composition to an epithelium of the target tissue, thecomposition comprising: a deconstructed matrisome including one or moreenzymatically fragmented peptides derived from at least one biologicaltissue, and one or more pharmaceutically acceptable or cosmeticallyacceptable excipients, wherein the deconstructed matrisome is in anamount of about 0.1% by weight to about 15% by weight of thecomposition; and wherein the one or more enzymatically fragmentedpeptides are configured to retain cell signaling ability, therebylowering the pH of the surface and reducing presence or growth of apathogen on the surface of the target tissue.

Embodiments of the invention are directed to a method of improving acharacteristic of a target skin tissue, the method comprising: topicallyadministering a composition to an epithelium of the target skin tissue,the composition comprising: a deconstructed matrisome including one ormore enzymatically fragmented peptides derived from at least onebiological tissue, and one or more pharmaceutically acceptable orcosmetically acceptable excipients, wherein the deconstructed matrisomeis in an amount of about 0.1% by weight to about 15% by weight of thecomposition; wherein the one or more enzymatically fragmented peptidesare configured to retain cell signaling ability, thereby improving thecharacteristic of the target skin tissue; and wherein the characteristicof the target skin tissue is selected from the group consisting offirmness, elasticity, fine lines, wrinkles, skin texture, skin tone, andappearance.

Embodiments of the invention are directed to a method of increasing cellregeneration in a target tissue, the method comprising: topicallyadministering a composition to an epithelium of the tissue, thecomposition comprising: a deconstructed matrisome including one or moreenzymatically fragmented peptides derived from at least one biologicaltissue, and one or more pharmaceutically acceptable or cosmeticallyacceptable excipients, wherein the deconstructed matrisome is in anamount of about 0.1% by weight to about 15% by weight of thecomposition; and wherein the one or more enzymatically fragmentedpeptides are configured to retain cell signaling ability, therebyrestoring an epithelial barrier of the target tissue.

BRIEF DESCRIPTION OF THE DRAWINGS

The accompanying drawings, which are incorporated in and form a part ofthe specification, illustrate the embodiments of the invention andtogether with the written description, serve to explain the principles,characteristics, and features of the invention. In the drawings:

FIG. 1 depicts an illustrative diagram of a method of making afibrosis-specific extracellular matrix substrate in accordance with anembodiment in accordance with an embodiment.

FIG. 2 depicts an example of normal human dermal fibroblasts cultured ondeconstructed matrisome in accordance with an embodiment.

FIG. 3 depicts an example of normal human dermal fibroblasts cultured ondeconstructed matrisome in accordance with an embodiment.

FIG. 4 depicts an example of an SDS-PAGE gel of blood vessel and skinextracellular matrix components in accordance with an embodiment.

FIG. 5 depicts an example of gene expression in extracellular matrixcomponents cultured with tissue-specific cells in accordance with anembodiment.

FIG. 6 depicts an example of biochemical analysis of blood vessel andskin extracellular matrix components in accordance with an embodiment.

FIG. 7 depicts an example of a matrikine-induced increase in dermaltissue regeneration in accordance with an embodiment.

FIG. 8 depicts an example of a matrikine-induced reduction in rednessduring wound healing in accordance with an embodiment.

FIG. 9 depicts an example of matrikine-induced reduction in wound sizein accordance with an embodiment.

FIG. 10 depicts an illustration of matrikine-induced healing inaccordance with an embodiment.

FIG. 11 depicts an example of matrikine-induced cellular migration andwound surface reduction in accordance with an embodiment.

FIGS. 12A and 12B depicts an example of a human repeat insult patch testin accordance with an embodiment.

FIG. 13 depicts an example of matrikine-induced wound healing inaccordance with an embodiment.

FIG. 14 depicts an example of matrikine-induced wound healing inaccordance with an embodiment.

FIG. 15 depicts an example of matrikine-induced wound healing inaccordance with an embodiment.

FIG. 16 depicts an example of matrikine-induced scar reduction inaccordance with an embodiment.

FIG. 17 depicts an example of matrikine-induced scar reduction inaccordance with an embodiment.

FIG. 18 depicts an example of matrikine-induced scar reduction inaccordance with an embodiment.

FIG. 19 depicts an example of matrikine-induced scar reduction inaccordance with an embodiment.

FIG. 20 depicts an example of matrikine-induced treatment of acnevulgaris in accordance with an embodiment.

FIG. 21 depicts an example of matrikine-induced treatment of acnevulgaris in accordance with an embodiment.

FIG. 22 depicts an example of matrikine-induced treatment of wrinkles inaccordance with an embodiment.

FIG. 23 depicts an illustration of user perception in accordance with anembodiment.

DETAILED DESCRIPTION

This disclosure is not limited to the particular systems, devices andmethods described, as these may vary. The terminology used in thedescription is for the purpose of describing the particular versions orembodiments only, and is not intended to limit the scope. Such aspectsof the disclosure be embodied in many different forms; rather, theseembodiments are provided so that this disclosure will be thorough andcomplete, and will fully convey its scope to those skilled in the art.

Various aspects will be described in detail hereinafter. Such aspectsmay, however, be embodied in many different forms and should not beconstrued as limited to the embodiments set forth herein; rather, theseembodiments are provided so that this disclosure will be thorough andcomplete, and will fully convey its scope to those skilled in the art.

The singular forms “a,” “an,” and “the” include plural referents unlessthe context clearly dictates otherwise. Thus, for example, reference toa “polymer” includes a single polymer as well as two or more of the sameor different polymers; reference to an “excipient” includes a singleexcipient as well as two or more of the same or different excipients,and the like.

The term “about,” as used herein, refers to variations in a numericalquantity that can occur, for example, through measuring or handlingprocedures in the real world; through inadvertent error in theseprocedures; through differences in the manufacture, source, or purity ofcompositions or reagents; and the like. Typically, the term “about” asused herein means greater or lesser than the value or range of valuesstated by 1/10 of the stated values, e.g., ±10%. The term “about” alsorefers to variations that would be recognized by one skilled in the artas being equivalent so long as such variations do not encompass knownvalues practiced by the prior art. Each value or range of valuespreceded by the term “about” is also intended to encompass theembodiment of the stated absolute value or range of values. Whether ornot modified by the term “about,” quantitative values recited in thepresent disclosure include equivalents to the recited values, e.g.,variations in the numerical quantity of such values that can occur, butwould be recognized to be equivalents by a person skilled in the art.Where the context of the disclosure indicates otherwise, or isinconsistent with such an interpretation, the above-statedinterpretation may be modified as would be readily apparent to a personskilled in the art. For example, in a list of numerical values such as“about 49, about 50, about 55, “about 50” means a range extending toless than half the interval(s) between the preceding and subsequentvalues, e.g., more than 49.5 to less than 52.5. Furthermore, the phrases“less than about” a value or “greater than about” a value should beunderstood in view of the definition of the term “about” providedherein.

The transitional term “comprising,” which is synonymous with“including,” “containing,” or “characterized by,” is inclusive oropen-ended and does not exclude additional, un-recited elements ormethod steps. By contrast, the transitional phrase “consisting of”excludes any element, step, or ingredient not specified in the claim.The transitional phrase “consisting essentially of” limits the scope ofa claim to the specified materials or steps “and those that do notmaterially affect the basic and novel characteristic(s)” of the claimedsubject matter. In some embodiments or claims where the term comprisingis used as the transition phrase, such embodiments can also beenvisioned with replacement of the term “comprising” with the terms“consisting of” or “consisting essentially of.”

It will be understood by those within the art that, in general, termsused herein are generally intended as “open” terms (for example, theterm “including” should be interpreted as “including but not limitedto,” the term “having” should be interpreted as “having at least,” theterm “includes” should be interpreted as “includes but is not limitedto,” et cetera). Further, the transitional term “comprising,” which issynonymous with “including,” “containing,” or “characterized by,” isinclusive or open-ended and does not exclude additional, unrecitedelements or method steps. While various compositions, methods, anddevices are described in terms of “comprising” various components orsteps (interpreted as meaning “including, but not limited to”), thecompositions, methods, and devices can also “consist essentially of” or“consist of” the various components and steps, and such terminologyshould be interpreted as defining essentially closed-member groups. Bycontrast, the transitional phrase “consisting of” excludes any element,step, or ingredient not specified in the claim. The transitional phrase“consisting essentially of” limits the scope of a claim to the specifiedmaterials or steps “and those that do not materially affect the basicand novel characteristic(s)” of the claimed invention.

All percentages, parts and ratios are based upon the total weight of theformulations and compositions and all measurements made are at about 25°C., unless otherwise specified.

As will be understood by one skilled in the art, for any and allpurposes, such as in terms of providing a written description, allranges disclosed herein are intended as encompassing each interveningvalue between the upper and lower limit of that range and any otherstated or intervening value in that stated range. All ranges disclosedherein also encompass any and all possible subranges and combinations ofsubranges thereof. Any listed range can be easily recognized assufficiently describing and enabling the same range being broken downinto at least equal halves, thirds, quarters, fifths, tenths, et cetera.As a non-limiting example, each range discussed herein can be readilybroken down into a lower third, middle third and upper third, et cetera.As will also be understood by one skilled in the art all language suchas “up to,” “at least,” and the like include the number recited andrefer to ranges that can be subsequently broken down into subranges asdiscussed above. Finally, as will be understood by one skilled in theart, a range includes each individual member. Thus, for example, a grouphaving 1-3 cells refers to groups having 1, 2, or 3 cells as well as therange of values greater than or equal to 1 cell and less than or equalto 3 cells. Similarly, a group having 1-5 cells refers to groups having1, 2, 3, 4, or 5 cells, as well as the range of values greater than orequal to 1 cell and less than or equal to 5 cells, and so forth.

In addition, even if a specific number is explicitly recited, thoseskilled in the art will recognize that such recitation should beinterpreted to mean at least the recited number (for example, the barerecitation of “two recitations,” without other modifiers, means at leasttwo recitations, or two or more recitations). Furthermore, in thoseinstances where a convention analogous to “at least one of A, B, and C,et cetera” is used, in general such a construction is intended in thesense one having skill in the art would understand the convention (forexample, “a system having at least one of A, B, and C” would include butnot be limited to systems that have A alone, B alone, C alone, A and Btogether, A and C together, B and C together, and/or A, B, and Ctogether, et cetera). In those instances where a convention analogous to“at least one of A, B, or C, et cetera” is used, in general such aconstruction is intended in the sense one having skill in the art wouldunderstand the convention (for example, “a system having at least one ofA, B, or C” would include but not be limited to systems that have Aalone, B alone, C alone, A and B together, A and C together, B and Ctogether, and/or A, B, and C together, et cetera). It will be furtherunderstood by those within the art that virtually any disjunctive wordand/or phrase presenting two or more alternative terms, whether in thedescription, sample embodiments, or drawings, should be understood tocontemplate the possibilities of including one of the terms, either ofthe terms, or both terms. For example, the phrase “A or B” will beunderstood to include the possibilities of “A” or “B” or “A and B.”

In addition, where features of the disclosure are described in terms ofMarkush groups, those skilled in the art will recognize that thedisclosure is also thereby described in terms of any individual memberor subgroup of members of the Markush group.

By hereby reserving the right to proviso out or exclude any individualmembers of any such group, including any sub-ranges or combinations ofsub-ranges within the group, that can be claimed according to a range orin any similar manner, less than the full measure of this disclosure canbe claimed for any reason. Further, by hereby reserving the right toproviso out or exclude any individual substituents, structures, orgroups thereof, or any members of a claimed group, less than the fullmeasure of this disclosure can be claimed for any reason.

The terms “patient” and “subject” are interchangeable and may be takento mean any living organism which may be administered and/or treatedwith compounds or compositions provided for herein. As such, the terms“patient” and “subject” may comprise, but is not limited to, anynon-human mammal, primate or human. In some embodiments, the “patient”or “subject” is a mammal, such as mice, rats, other rodents, rabbits,dogs, cats, swine, cattle, sheep, horses, primates, or humans. In someembodiments, the patient or subject is an adult, child, or infant. Insome embodiments, the patient or subject is a human.

Generally speaking, the term “tissue” refers to any aggregation ofsimilarly specialized cells which are united in the performance of aparticular function. As used herein, “tissue”, unless otherwiseindicated, refers to tissue which includes elastin as part of itsnecessary structure and/or function. For example, connective tissuewhich is made up of, among other things, collagen fibrils and elastinfibrils satisfies the definition of “tissue” as used herein.Additionally, elastin appears to be involved in the proper function ofblood vessels, veins, and arteries in their inherent visco-elasticity.

The term “composition” as used herein refers to a combination or amixture of two or more different ingredients, components, or substances.

The term “matrikine” as used herein refers to extracellularmatrix-derived peptides which regulate cell activity. Matrikines arebioactive peptides generated as a result of partial enzymaticdegradation (i.e., proteolysis) of extracellular matrix macromolecules.The peptides are fragments of the full-length molecules and oftenmodulate cellular activities through signaling in different manners thanthe full-length molecules, including but not limited to modulating cellproliferation, migration, and apoptosis. Furthermore, the signaling frommatrikines may vary based the degree of fragmentation that occursthrough enzymatic degradation (i.e. the size of the fragments). The term“Matrikyne” as used here refers to the formulations of matrikinesdisclosed in the present application and/or produced by XYLYX BIO, INC.of New York, USA.

The term “matrisome” as used herein refers to the set of matrikinesreleased from an extracellular matrix or resulting from the enzymaticdegradation of an extracellular matrix. A complete matrisome maycomprise different concentrations of matrikines depending on thetissue-specific extracellular matrix the matrisome is derived from andrepresents a full complement of matrikines.

The term “excipients” as used herein encompasses carriers and diluents,meaning a material, composition or vehicle, such as a liquid or solidfiller, diluent, excipient, solvent or encapsulating material involvedin carrying or transporting a pharmaceutical, cosmetic or other agentacross a tissue layer such as the stratum corneum or stratum spinosum.

The term “keratinous fiber” as used herein refers to any tissue whichcontain keratin as a fibrous structural protein, including, but notlimited to, skin, hair, and nails.

The terms “topically” and “topical” as used herein refer to applicationof the compositions to the surface of the skin, mucosal cells, keratinsand tissues. Examples of keratins are nails and hair.

The phrase “pharmaceutically acceptable” or “cosmetically acceptable” isemployed herein to refer to those agents of interest/compounds, salts,compositions, dosage forms, etc., which are within the scope of soundmedical judgment suitable for use in contact with the tissues of humanbeings and/or other mammals without excessive toxicity, irritation,allergic response, or other problem or complication, commensurate with areasonable benefit/risk ratio. In some aspects, pharmaceuticallyacceptable means approved by a regulatory agency of the federal or astate government, or listed in the U.S. Pharmacopeia or other generallyrecognized pharmacopeia for use in mammals (e.g., animals), and moreparticularly, in humans.

The term “cosmetic” means an agent utilized, and/or intended to beapplied to the human body for cleansing, beautifying, promotingattractiveness, altering the appearance of the skin or any combinationthereof.

As used herein, the term “therapeutic” means an agent utilized to treat,combat, ameliorate, prevent, or improve an unwanted condition or diseaseof a patient. In part, embodiments of the present invention are directedto the treatment of wounds, scars, and skin conditions.

In some embodiments, the compounds and methods disclosed herein can beutilized with or on a subject in need of such treatment, which can alsobe referred to as “in need thereof” As used herein, the phrase “in needthereof” means that the subject has been identified as having a need forthe particular method or treatment and that the treatment has been givento the subject for that particular purpose.

The term “treat,” “treated,” or “treating” as used herein, refers tomethods of treating a skin disorder or a systemic condition, andgenerally includes the administration of a compound or composition,wherein the object is to reduce the frequency of, or delay the onset of,symptoms of a medical condition, enhance the texture, appearance, color,sensation, or hydration of the intended tissue treatment area of thetissue surface in a subject relative to a subject not receiving thecompound or composition, or to otherwise obtain beneficial or desiredclinical results. For the purposes of this invention, beneficial ordesired clinical results include, but are not limited to, reversal,reduction, or alleviation of symptoms of a condition; diminishment ofthe extent of the condition, disorder or disease; stabilization (i.e.,not worsening) of the state of the condition, disorder or disease; delayin onset or slowing of the progression of the condition, disorder ordisease; amelioration of the condition, disorder or disease state; andremission (whether partial or total), whether detectable orundetectable, or enhancement or improvement of the condition, disorderor disease. Treatment includes eliciting a clinically significantresponse without excessive levels of side effects. Treatment alsoincludes prolonging survival as compared to expected survival if notreceiving treatment.

The term “inhibiting” includes the administration of a compound of thepresent invention to prevent the onset of the symptoms, alleviating thesymptoms, reducing the symptoms, delaying or decreasing the progressionof the disease and/or its symptoms, or eliminating the disease,condition or disorder.

For convenience, certain terms employed in the specification, examplesand claims are collected here. Unless defined otherwise, all technicaland scientific terms used in this disclosure have the same meanings ascommonly understood by one of ordinary skill in the art to which thisdisclosure belongs.

Throughout this disclosure, various patents, patent applications andpublications are referenced. The disclosures of these patents, patentapplications and publications in their entireties are incorporated intothis disclosure by reference in order to more fully describe the stateof the art as known to those skilled therein as of the date of thisdisclosure. This disclosure will govern in the instance that there isany inconsistency between the patents, patent applications andpublications cited and this disclosure.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meanings as commonly understood by one of ordinary skillin the art. Nothing in this disclosure is to be construed as anadmission that the embodiments described in this disclosure are notentitled to antedate such disclosure by virtue of prior invention.

As discussed herein, pro-regeneration biomaterial peptides derived fromextracellular matrix (i.e., matrikines) can provide cells with thecomplex structural support and biochemical signals required for cellularregeneration. Bioactive compositions may provide comprehensive skinrepair and overall skin health using regenerative bioactive ingredients.The compositions may be varied to create a product line that includesproducts aimed at scar treatment and other skin conditions, such asacne, eczema, and psoriasis (e.g., Matrikyne® compositions availablefrom XYLYX BIO, INC.).

Hundreds of matrikine bioactive protein complexes promote skin cellregeneration and healing, thereby providing a clinically proveningredient with proven rejuvenating properties. The bioactive componentsorchestrate essential communication between skin cells, the healthy skinmicrobiome and the immune system, integrally supporting active skin cellregeneration and health. The hundreds of matrikine bioactive proteincomplexes come from six broad families of bioactive molecules that worktogether to contribute essential repair functions. Bioactive familiesand their benefits include: laminins (regulate cell migration anddifferentiation), proteoglycans (signal skin cell renewal andproliferation), growth factors (support skin cell rejuvenation),glycoproteins (direct immune cells and system), elastin (support intissue integrity and tone), collagens (support skin repair andregeneration).

Natural matrikine bioactivity supports tissue repair and healing byregulating inflammation, inhibiting fibrosis and enhancing production ofcollagen and expression of growth factors. Matrikines guide cellularregeneration and are integral to physiological ‘dialogue’ underlyingskin health. Unlike conventionally available products, fragmentedextracellular matrix includes matrikines and is able to penetrate theskin to facilitate dialogue between skin cells, the immune system andthe microbiome. Formulations can be targeted to various applications,including reduction in the appearance of fine lines and wrinkles, scarand acne treatment, and overall skin brightening, firming andrejuvenation. Matrikines are also proven to reduce redness by restoringthe skin's natural balance and can be used for scar treatment, rednessreduction, eczema, roseacea and as an anti-wrinkle treatment.Furthermore, matrikines have the ability to tailor to skin condition,age ethnicity, etc. for product personalization.

Topical Matrikine Compositions

Embodiments disclosed herein are directed to a composition for topicalapplication comprising fragmented deconstructed matrisome including oneor more peptides (i.e., matrikines). The composition may furthercomprise one or more excipients (e.g., a pharmaceutically acceptableexcipient or a cosmetically acceptable excipient). In some embodiments,the deconstructed matrisome may be fragmented to generate the one ormore peptides through proteolysis (i.e., enzymatic degradation) of oneor more deconstructed matrisome components. Further, the one or morepeptides may be configured to promote one or more of tissue homeostasis,tissue repair, and tissue regeneration.

The matrikines may be may be derived from tissue-specific extracellularmatrix (TS-ECM) specific to a variety of tissue types, and thus theresulting mixture of matrikines in the composition may emulate the nicheenvironment of various tissues. In some embodiments, the TS-ECM maycomprise skin-specific ECM. However, a variety of TS-ECMs may be used toderive the matrikines. In some embodiments, the TS-ECM may be selectedfrom blood vessel-specific ECM, cartilage-specific ECM,intestine-specific ECM, liver-specific ECM, placenta-specific ECM,skin-specific ECM and stomach-specific ECM. In still additionalembodiments, the TS-ECM may emulate a niche environment specific toanother tissue. For example, the tissue may be selected from the adrenalgland, amnion, bladder, bone, brain, breast, chorion, connective tissue,esophagus, eye, fat, heart, kidney, larynx, ligament, lung, lymph,microvasculature, muscle, mouth, omentum, ovary, fallopian tube,thyroid, parathyroid, large intestine, small intestine, pancreas,peritoneum, pharynx, placenta membrane, prostate, rectum, smooth muscle,spinal cord, spinal fluid, spleen, tendon, testes, thymus, umbilicalcord, uterus, vagina, or Wharton's Jelly. In some embodiments, theTS-ECM may emulate a region of the anatomy, an organ, or a region of anorgan. For example, a TS-ECM may represent the large intestine or it maymore specifically represent the colon or the rectum.

In some embodiments, the deconstructed matrisome may be derived frombiological tissue. A variety of tissue sources may be used as a startingmaterial to obtain deconstructed matrisome. In some embodiments, thetissue source is a human source. In some embodiments, the tissue sourceis an animal source. For example, the tissue may be porcine (i.e.,sourced from a pig) or any other animal tissue known to have clinicalrelevance.

In some embodiments, the tissue source may be other than a human oranimal source. In some embodiments, the tissue source may be selectedfrom a plant source and a fungal source. While the deconstructedmatrisomes exemplified herein are representative of animal tissues, theselected tissue may vary based on the type of organism selected as atissue source. For example, where the tissue source is a plant sourceand/or a fungal source, the selected tissue may be any tissue typesknown to be present in plants and/or fungi as understood by a personhaving an ordinary level of skill in the art.

In some embodiments, the tissue source is selected from fetal tissue,juvenile tissue, and adult tissue. In some embodiments, the tissuesource is healthy tissue. However, in some embodiments, the tissuesource is selected from diseased tissue, transgenic tissue, or tissuehaving a specific disorder or health condition. For example, specificdisorders or conditions may result in overexpression of specificproteins and/or peptides. Accordingly, it may be beneficial to apply acomposition comprising the specific proteins or matrikines to anindividual exhibiting under expression of the same specific proteins ormatrikines. Therefore, it may be advantageous to derive the matrikinesfrom TS-ECM from a non-healthy tissue in a targeted manner. Theresulting TS-ECM is representative of extracellular matrix from thetissue source, or more generally from tissue having the same relevantcharacteristics as the tissue source (e.g., fetal human skin tissue willyield skin-specific ECM representative of fetal human skin).

In some embodiments, the matrikines are isolated from a single TS-ECM.In some embodiments, the matrikines are isolated from a plurality ofTS-ECMs. Accordingly, the mixture of matrikines and relativeconcentrations of matrikines included in the composition may represent asingle tissue a combination of tissues. The plurality of TS-ECMs may beprovided in predetermined quantities or ratios with respect to oneanother in order to tailor the resulting mixture of matrikines. Further,any number of matrikines isolated from the TS-ECM may be adjusted (e.g.,adjusting a concentration) or entirely removed in order to produce acomposition having a matrikine mixture with desired properties fortissue repair and regeneration.

In some embodiments, the composition includes enzymatically fragmenteddeconstructed matrisome and/or the matrikines in specifiedconcentrations that emulates the extracellular environment found in aspecific native tissue. The composition may include a specificcombination of deconstructed matrisome macromolecules such asscaffolding proteins, ECM-associated proteins, ECM regulators,glycoproteins, proteoglycans, laminins, extracellular matrix associateproteins, soluble growth factors, inflammatory cytokines and chemokines,immune mediators, and secreted factors in the extracellular fluid.Additionally, the composition may include a specific combination ofmatrikines. Because matrikines are derived from the macromolecules inthe TS-ECM (i.e., ‘parent’ proteins or macromolecules), the mixture ofmatrikines (i.e., ‘child’ peptides) in the composition may varyaccording to the makeup of the TS-ECM. Therefore, the matrikines in thecomposition may correspond to the type of TS-ECM from which thematrikines are sourced. Further, each of these components may havesubtypes, the presence of each of which may vary from one deconstructedmatrisome to another deconstructed matrisome. Each deconstructedmatrisome may be characterized by the presence or absence of one or morecomponents. Further, the concentration of each component may vary fromone deconstructed matrisome to another deconstructed matrisome.

As described above, ECM comprises macromolecules (e.g., proteins.lipids, and polysaccharides) and other factors that are specific forcell-signaling in a particular niche-environment. In native ECM, the ECMcomponents form a three-dimensional ultrastructure. In someapplications, one may prefer a more uniform substrate like a solution ora hydrogel, for example, for topical application. The TS-ECM produced bythe methods described herein is distinct from native ECM. The TS-ECM isdecellularized and the removal of the cellular structure modulates theconcentrations of macromolecules and other cell-signaling factors.Further, the three-dimensional ultrastructure may be removed and thevarious components of the deconstructed matrisome may be digested intofragments. Any of the ECM components in the various embodiments and/orformulas described herein may be fragmented in the deconstructedmatrisome, including but not limited to collagen, elastin,glycosaminoglycans, proteoglycans, matrix associated factors, ECMregulators, matrisome secreted factors, immune factors, marrowassociated factors, and other structural factors. The removal of thethree-dimensional ultrastructure of the ECM and the fragmentation ofdeconstructed matrisome components facilitates formation of a homogenousmixture for use in forming compositions for topical application, e.g.,hydrogels, liquid solution, powders, and other formats as describedherein. Surprisingly, the fragmented components nonetheless contributeto cell signaling along with small molecules, thus retaining thecharacteristics of the niche environment to a high degree despite thefragmentation and lack of ultrastructure which are needed to form theconventional substrate structure.

In some embodiments, the deconstructed matrisome comprises a homogenousmixture of macromolecule fragments including collagen, elastin, andglycosaminoglycan. In some embodiments, the deconstructed matrisomecomprises macromolecules or macromolecule fragments including collagen,elastin, and glycosaminoglycan, wherein the amount of each macromoleculemay be decreased after decellularization. In some embodiments, thedeconstructed matrisome comprises a homogenous mixture of macromoleculefragments including collagen, elastin, and glycosaminoglycan, whereinthe concentration of each macromolecule may be changed afterdecellularization.

In some embodiments, the deconstructed matrisome comprises a homogenousmixture of macromolecule fragments. In some embodiments, thedeconstructed matrisome comprises a homogenous mixture of macromoleculefragments, wherein the macromolecules may be fully or partiallyfragmented after enzymatic digestion. In some embodiments, thedeconstructed matrisome comprises a homogenous mixture of macromoleculefragments, wherein the homogenous mixture retains cellular signaling. Insome embodiments, the deconstructed matrisome comprises a homogenousmixture of macromolecule fragments, wherein the homogenous mixture doesnot contain the ECM three-dimensional ultra-structure. In someembodiments, the ECM three-dimensional ultra-structure is not requiredfor cell-matrix recognition. In some embodiments, interactionsresponsible for cell-matrix recognition is not limited to structuralcues from acellular matrix, but also relies on signaling from smallmolecules or protein fragments. In some embodiments described herein,the deconstructed matrisome is processed into an ECM powder. In someembodiments, the ECM powder comprises a homogenous mixture ofmacromolecule fragments, wherein the macromolecules may be fully orpartially fragmented after enzymatic digestion. In some embodiments, theECM powder comprises a homogenous mixture of macromolecule fragments,wherein the homogenous mixture retains cellular signaling. In someembodiments, the ECM powder comprises a homogenous mixture ofmacromolecule fragments, wherein the homogenous mixture does not containthe ECM three-dimensional ultra-structure. In some embodiments, the ECMthree-dimensional ultra-structure is not required for cell-matrixrecognition. In some embodiments, interactions responsible forcell-matrix recognition is not limited to structural cue fromdecellularized matrix, but also relies on signaling from small moleculesor protein fragments.

In some embodiments, the TS-ECM may not be enzymatically digested andthe three-dimensional ultrastructure may be maintained, e.g., as anacellular and/or dehydrated scaffold.

The composition may be characterized by any matrikine components,concentrations thereof, and/or changes thereof from normal as summarizedin Table 1, Table 2, and Table 3, Table 4, Table 5, Table 6, and Table7. While the Tables describe various formulas with respect to the parentECM macromolecules detected therein, it is understood that thecomposition may include the matrikines, or fragmented peptides, derivedfrom the corresponding parent ECM macromolecules. However, thesecompositions are exemplary in nature and the matrikine profiles may varytherefrom as to any number of components. For example, the compositionmay vary from the described concentration values and/or ranges by about10%, about 20%, about 30%, greater than 30%, or individual values orranges therebetween.

In some embodiments, the deconstructed matrisome comprises collagen. Insome embodiments, the deconstructed matrisome comprises fragmentedcollagen. In some embodiments, the deconstructed matrisome comprisescollagen in an amount from about 400 μg/mL to about 9700 μg/mL. In someembodiments, the deconstructed matrisome comprises collagen in an amountfrom about 2900 μg/mL to about 3800 μg/mL. In some embodiments, thedeconstructed matrisome comprises collagen in an amount from about 7500μg/mL to about 9500 μg/mL. In some embodiments, the deconstructedmatrisome comprises collagen from about 1100 μg/mL to about 1300 μg/mL.In some embodiments, the deconstructed matrisome comprises collagen fromabout 400 μg/mL to about 530 μg/mL. In some embodiments, thedeconstructed matrisome comprises collagen from about 7000 μg/mL toabout 9700 μg/mL. In some embodiments the collagen is collagen type IV,wherein the collagen type IV is in an amount of about 2 ng/mL to about24 ng/mL. In some embodiments the collagen is collagen type IV, whereinthe collagen type IV is in an amount of about 6 ng/mL to about 10 ng/mL.In some embodiments the collagen is collagen type IV, wherein thecollagen type IV is in an amount of about 20 ng/mL to about 24 ng/mL. Insome embodiments the collagen is collagen type IV, wherein the collagentype IV is in an amount of about 9 ng/mL to about 19 ng/mL. In someembodiments the collagen is collagen type IV, wherein the collagen typeIV is in an amount of about 2 ng/mL to about 10 ng/mL. In someembodiments the collagen is collagen type IV, wherein the collagen typeIV is in an amount of about 3 ng/mL to about 5 ng/mL.

In some embodiments, the deconstructed matrisome comprises elastin. Insome embodiments, the deconstructed matrisome comprises fragmentedelastin. In some embodiments, the deconstructed matrisome compriseselastin in an amount from about 40 μg/mL to about 3000 μg/mL. In someembodiments the elastin is in an amount from about 40 μg/mL to about 50μg/mL. In some embodiment the elastin is in an amount from about 350μg/mL to about 450 μg/mL. In some embodiments the elastin is in anamount from about 120 μg/mL to about 150 μg/mL. In some embodiments theelastin is in an amount from about 1700 μg/mL to about 3000 μg/mL.

In some embodiments, the deconstructed matrisome comprisesglycosaminoglycan. In some embodiments, the deconstructed matrisomecomprises fragmented glycosaminoglycan. In some embodiments, thedeconstructed matrisome comprises glycosaminoglycan in an amount fromabout 3 μg/ml to about 170 μg/ml. in some embodiments, theglycosaminoglycan is in an amount from about 130 μg/mL to about 170μg/mL. In some embodiments, the glycosaminoglycan is in an amount fromabout 10 μg/mL to about 20 μg/mL. In some embodiments, theglycosaminoglycan is in an amount from about 5 μg/mL to about 15 μg/mL.In some embodiments, the glycosaminoglycan is in an amount from about 3μg/mL to about 5 μg/mL. In some embodiments, the glycosaminoglycan is inan amount from about 80 μg/mL to about 110 μg/mL.

TABLE 1 Partial list of components in Matrikynes Formula 1. MATRIKYNES ™FORMULA 1 Protein Category Description Collagens Type I, alpha 1 chainType III, alpha 1 chain Type V, alpha 2 chain Glycoproteins Fibrillarcollagen NC1 domain-containing protein Fibrillin 1 Microfibrilassociated protein 4 Proteoglycan Heparan sulfate proteoglycan 2 ElastinElastin isoform Structural proteins Actin gamma 2 Filamin A Growthfactors Latent transforming growth factor beta binding protein 4

TABLE 2 Partial list of components in Matrikynes Formula 2. MATRIKYNES ™FORMULA 2 Protein Category Description Collagens Type I, alpha 1 chainType I, alpha 2 chain Type II, alpha 1 chain Type III, alpha 1 chainType V, alpha 1 chain Type V, alpha 2 chain Type VI, alpha 2 chain TypeVI, alpha 3 chain Type VIII, alpha 1 chain Type IX, alpha 2 chain TypeXI, alpha 1 chain Type XI, alpha 2 chain Type XII, alpha 2 chain TypeXIV, alpha 1 chain Glycoproteins Adipocyte enhancer binding protein 1Alpha-2-Heremans-Schmid glycoprotein Biglycan Extracellular matrixprotein 2 Fibrillin 1 Fibrinogen beta chain Fibrinogen gamma chainFibronectin 1 Osteonectin Periostin Tenascin C Tenascin N Thrombospondin1 Transforming growth factor beta induced Vitronectin ProteoglycansAggrecan core protein Asporin Decorin Fibromodulin Heparan sulfateproteoglycan 2 Lumican Mimecan Osteoglycan OsteomodulinProline/arginine-rich end leucine-rich repeat protein Elastin ElastinMatrisome secreted Albumin factors Annexin A2 Chitinase Collectinsubfamily member 12 Creatine kinase B Olfactomedin ECM regulatorsCoagulation factor IX Coagulation factor X Inter-alpha (globulin)inhibitor H4 Prothrombin Serpin peptidase inhibitor clade F Structuralproteins Actin gamma 2 Vimentin

TABLE 3 Partial list of components in Matrikynes Formula 3. MATRIKYNES ™FORMULA 3 Protein Category Description Collagens Type I, alpha 1 chainType I, alpha 2 chain Type II, alpha 1 chain Type III, alpha 1 chainType IV, alpha 1 chain Type IV, alpha 2 chain Type V, alpha 1 chain TypeV, alpha 2 chain Type VI, alpha 2 chain Type VI, alpha 3 chain Type VI,alpha 5 chain Type VIII, alpha 1 chain Type VIII, alpha 2 chainGlycoproteins Dermatopontin Fibrillin 1 Microfibril-associated protein 4Periostin Proteoglycans Asporin Heparan sulfate proteoglycan 2 ElastinElastin isoform Matrisome secreted Chitinase factors Collectin subfamilymember Trefoil factor 1 Vasoactive intestinal peptide ECM regulatorsHyaluronan binding protein 2 Structural Proteins Actin gamma 2 Myosin 11Growth Factors Amphiregulin Basic fibroblast growth factor Bonemorphogenic protein 4 Bone morphogenic protein 7 Epidermal growth factorGrowth differentiation factor 15 Hepatocyte growth factor Insulin-likegrowth factor binding protein 3 Osteoprotegerin

TABLE 4 Partial list of components in Matrikynes Formula 4. MATRIKYNES ™FORMULA 4 Protein Category Description Collagens Type I, alpha 1 chainType I, alpha 2 chain Type II, alpha 1 chain Type III, alpha 1 chainType IV, alpha 1 chain Type V, alpha 2 chain Type VI, alpha 3 chain TypeVI, alpha 5 chain Glycoproteins EGF-containing fibulin-likeextracellular matrix protein Fibrillin 1 Fibrillin 2 Laminin subunitgamma 1 Prostate stem cell antigen Saposin-B-Val Von Willebrand factorProteoglycans Heparan sulfate proteoglycan Elastin Elastin isoformMatrisome secreted Chitinase factors Mucin 5AC Mucin 6 Serum albuminTrefoil factor 2 ECM regulators Granulin precursor Structural proteinsActin Keratin 1 Keratin 2 Keratin 9 Keratin 10 Myosin heavy chain 9Tubulin beta chain Growth Factors Bone morphogenic protein 4 Fibroblastgrowth factor 2 Insulin-like growth factor binding protein 4 Macrophagecolony-stimulating factor 1 receptor (CD115) Pro-epidermal growth factor

TABLE 5 Partial list of components in Matrikynes Formula 5. MATRIKYNES ™FORMULA 5 Protein Category Description Collagens Type I, alpha 1 chainType I, alpha 2 chain Type I, alpha 3 chain Type II, alpha 1 chain TypeIV, alpha 1 chain Type IV, alpha 2 chain Type IV, alpha 3 chain Type IV,alpha 4 chain Type IV, alpha 5 chain Type V, alpha 1 chain Type V, alpha2 chain Type VI, alpha 1 chain Type VI, alpha 2 chain Type VI, alpha 3chain Type VIII, alpha 1 chain Type XVI, alpha 1 chain Type XXI, alpha 1chain Glycoproteins Dermatopontin Fibulin 2 Laminin subunit alpha 3Laminin subunit alpha 5 Laminin subunit beta 2 Laminin subunit gamma 1Nidogen 1 Periostin Vitronectin Proteoglycans Biglycan Heparan sulfateproteoglycan core protein Elastin Elastin isoform Matrisome secretedHornerin factors ECM regulators Alpha 1 antitrypsin Cathepsin GDesmoplakin Junction plakoglobin Serum albumin precursorMetalloproteinase Inhibitor 3 Structural proteins Keratin 1 Keratin 2Keratin 5 Keratin 9 Keratin 10 Keratin 14 Growth Factors Basicfibroblast growth factor Brain-derived neurotrophic factor Endocrinegland-derived vascular endothelial growth factor Epidermal growth factorreceptor Growth differentiation factor 15 Hepatocyte growth factorInsulin-like growth factor binding protein 1 Insulin-like growth factorbinding protein 6 Osteoprotegerin Platelet-derived growth factor AAVascular endothelial growth factor

TABLE 6 Quantification of extracellular matrix components in fiveMatrikynes ™ Formulas. ECM Component Formula 1 Formula 2 Formula 3Formula 4 Formula 5 Collagen type IV 6-10 20-24 9-19 2-10 3-5 (ng/mL)

TABLE 7 Major matricryptins/matrikines released from human extracellularparent proteins Matrikines Released from Human ECM Proteins MatrikineMatrikine Molecular Mass Parent Protein Common Name (kDa) Aggrecan coreprotein + Metastatin (hyaluronan-binding 85 + 38 Proteoglycan linkprotein complex) Collagen IV alpha 1 chain Arresten 26 Collagen IV alpha2 chain Canstatin 24 Collagen IV alpha 3 chain Tumstatin 27 Collagen IValpha 4 chain Tetrastatin 2.1-25 Collagen IV alpha 5 chain Pentastatin 12.5 Pentastatin 2 2.4 Pentastatin 3 2.1 Lamstatin 25 Collagen IV alpha 6chain Hexastatin 1 2 Heastatin 2 2.5 Collagen VI alpha 6 chainEndotrophin 5.8 (collagen-like 5 domain) Collagen XIII alpha 1 chainEctodomain of collagen XII 240 Collagen XV alpha 1 chain Restin 1 21Restin 2 20 Restin 3 19.4 Restin 4 18.4 Collagen XVII alpha 1 chainEctodomain of collagen XVII 120 Collagen XVIII alpha 1 chain Endostatin21 Neostatin 7 28 Neostatin 14 23 Collagen XIX alpha 1 chain NC1 domainof collagen XIX 2.2 Collagen XXIII alpha 1 chain Ectodomain of collagenXXIII 180 Collagen XXV alpha 1 chain Ectodomain of collagen XXV 158(collagen-like Alzheimer amyloid plaque component) FibronectinAnastellin 10 Fibstatin 29 Laminins Laminin peptides MMP-2 PEX 24Perlecan Endorepellin 85 Procollagen C-proteinase CUB1CUB2 domain 30enhancer 1 Tenascin-C Ten1/2 10 Ten11/12/13 3.4 Ten14 Syndecan-1Ectodomain of syndecan-1 24 Syndecan-2 Ectodomain of syndecan-2 14Syndecan-3 Ectodomain of syndecan-3 39 Syndecan-4 Ectodomain ofsyndecan-4 14 Tropoelastin Elastokines 0.5-8 

In some embodiments the composition comprises one or more of collagen,glycoprotein, proteoglycan, elastin, matrisome secreted factors,structural proteins, growth factors, and ECM regulators.

In some embodiments the composition comprises one or more fragments ofcollagen, glycoprotein, proteoglycan, elastin, matrisome secretedfactors, structural proteins, growth factors, and ECM regulators.

In some embodiments the compositions comprises collagen or fragments ofcollagen, wherein the collagen is selected from one or more of collagentype I alpha 1 chain, collagen type I alpha 2 chain, collagen type Ialpha 3 chain, collagen type II alpha 1 chain, collagen type III alpha 1chain, collagen type IV alpha 1 chain, collagen type IV alpha 2 chain,collagen type IV alpha 3 chain, collagen type IV alpha 4 chain, collagentype IV alpha 5 chain, collagen type V alpha 1 chain, collagen type Valpha 2 chain, collagen type VI alpha 1 chain, collagen type VI alpha 2chain, collagen type VI alpha 3 chain, collagen type VI alpha 5 chain,collagen type VIII alpha 1 chain, collagen type VIII alpha 2 chain,collagen type IX alpha 2 chain, collagen type XI alpha 1 chain, collagentype XI alpha 2 chain, collagen type XII alpha 2 chain, collagen typeXIV alpha 1 chain, collagen type XVI alpha 1 chain, collagen type XXIalpha 1 chain.

In some embodiments the composition comprises glycoproteins or fragmentsof glycoproteins, wherein the glycoprotein is selected from one or moreof fibrillar collagen NC1 domain-containing protein, fibrillin,fibrillin 1, fibrillin 2, fibulin 2, microfibril associated protein 4,adipocyte enhancer binding protein 1, alpha-2-Heremans-Schmidglycoprotein, biglycan, extracellular matrix protein 2, fibrinogen betachain, fibrinogen gamma chain, fibronectin 1, osteonectin, periostin,tenascin C, tenascin N, thrombospondin 1, transforming growth factorbeta induced, vitronectin, dermatopontin, EGF-containing fibulin-likeextracellular matrix protein, laminin, subunit alpha 3, laminin subunitealpha 5, laminin subunit beta 2, laminin subunit gamma 1, prostate stemcell antigen, saposin-B-Val, von Willebrand factor, and nidogen 1.

In some embodiments the composition comprises proteoglycans or fragmentsof proteoglycans, wherein the proteoglycan is selected from one or moreof heparan sulfate proteoglycan, heparan sulfate proteoglycan 2, heparansulfate proteoglycan core protein, aggrecan core protein, asporin,decorin, fibromodulin, lumican, mimecan, osteoglycan, osteomodulin,proline/arginine-rich end leucine-rich repeat protein, and biglycan.

In some embodiments, the composition comprises elastin or fragments ofelastin, wherein the elastin is elastin isoform.

In some embodiments, the composition comprises matrisome secretedfactors or fragments of matrisome secreted factors, wherein thematrisome secreted factors are selected from one or more of albumin,serum albumin, annexin A2, chitinase, collectin subfamily member 12,creatine kinase B, olfactomedin, trefoil factor 1, trefoil factor 2,vasoactive intestinal peptide, mucin SAC, mucin 5, and hornerin.

In some embodiments, the composition comprises structural proteins orfragments of structural proteins, wherein the structural proteins areselected from one or more of actin, actin gamma 2, filamin A, keratin 1,keratin 2, keratin 5, keratin 9, keratin 10, keratin 14, myosin 11,myosin heavy chain 9, tubulin beta chain, and vimentin.

In some embodiments, the composition comprises growth factors orfragments of growth factors, wherein the growth factors are selectedfrom one or more of latent transforming growth factor beta bindingprotein 4, amphiregulin, basic fibroblast growth factor, bonemorphogenic protein 4, bone morphogenic protein 7, brain-derivedneurotrophic factor, epidermal growth factor, epidermal growth factorreceptor, pro-epidermal growth factor, endocrine gland-derived vascularendothelial growth factor, fibroblast growth factor 2, growthdifferentiation factor 15, hepatocyte growth factor, insulin-like growthfactor binding protein 1, insulin-like growth factor binding protein 3,insulin-like growth factor binding protein 4, insulin-like growth factorbinding protein 6, macrophage colony-stimulating factor 1 receptor(CD115), osteoprotegerin, platelet-derived growth factor AA, vascularendothelial growth factor.

In some embodiments, the composition comprises ECM regulators orfragments of ECM regulators, wherein the ECM regulators are selectedfrom one or more of alpha 1 antitrypsin, cathepsin G, coagulation factorIX, coagulation factor X, desmoplakin, granulin precursor, hyaluronanbinding protein 2, inter-alpha (globulin) inhibitor H4, junctionplakoglobin, metalloproteinase inhibitor 2, prothrombin, serpinpeptidase inhibitor Glade F, and serum albumin precursor.

In some embodiments, the deconstructed matrisome comprises one or morefragments of collagens, glycoproteins, proteoglycans, elastins,matrisome secreted factors, structural proteins, growth factors, and ECMregulators, wherein the one or more collagens comprise collagen type Ialpha 1 chain, collagen type III alpha 1 chain, and collagen type Valpha 2 chain; wherein one or more glycoproteins comprise fibrillarcollagen NC1 domain-containing protein, fibrillin 1, and microfibrilassociated protein 4; wherein the one or more proteoglycans compriseheparan sulfate proteoglycan 2; wherein the one or more elastinscomprise elastin isoform; wherein the one or more structural proteinscomprise actin gamma 2 and filamin A; and wherein the one or more growthfactors comprise latent transforming growth factor beta binding protein4.

In some embodiments, the deconstructed matrisome comprises one or morefragments of collagens, glycoproteins, proteoglycans, elastins,matrisome secreted factors, structural proteins, growth factors, and ECMregulators, wherein the one or more collagens comprise collagen type Ialpha 1 chain, collagen type I alpha 2 chain, collagen type II alpha 1chain, collagen type III alpha 1 chain, collagen type V alpha 1 chain,collagen type V alpha 2 chain, collagen type VI alpha 2 chain, collagentype VI alpha 3 chain, collagen type VIII alpha 1 chain, collagen typeIX alpha 2 chain, collagen type XI alpha 1 chain, collagen type XI alpha2 chain, collagen type XII alpha 2 chain, and collagen type XIV alpha 1chain; wherein the one or more glycoproteins comprise fibrillin 1,adipocyte enhancer binding protein 1, alpha-2-Heremans-Schmidglycoprotein, biglycan, extracellular matrix protein 2, fibrinogen betachain, fibrinogen gamma chain, fibronectin 1, osteonectin, periostin,tenascin C, tenascin N, thrombospondin 1, transforming growth factorbeta induced, and vitronectin; wherein the one or more proteoglycanscomprise heparan sulfate proteoglycan 2, aggrecan core protein, asporin,decorin, fibromodulin, lumican, mimecan, osteoglycan, osteomodulin, andproline/arginine-rich end leucine-rich repeat protein; wherein the oneor more elastins comprise elastin; wherein the one or more mastrisomesecreted factors comprise albumin, annexin A2, chitinase, collectinsubfamily member 12, creatine kinase B, olfactomedin; wherein the one ormore ECM regulators are coagulation factor IX, coagulation factor X,inter-alpha (globulin) inhibitor H4, prothrombin, and serpin peptidaseinhibitor Glade F; and wherein the one or more structural proteins areactin gamma 2 and vimentin.

In some embodiments, the deconstructed matrisome comprises one or morefragments of collagens, glycoproteins, proteoglycans, elastins,matrisome secreted factors, structural proteins, growth factors, and ECMregulators, wherein the one or more collagens comprise collagen type Ialpha 1 chain, collagen type I alpha 2 chain, collagen type II alpha 1chain, collagen type III alpha 1 chain, collagen type IV alpha 1 chain,collagen type IV alpha 2 chain, collagen type V alpha 1 chain, collagentype V alpha 2 chain, collagen type VI alpha 2 chain, collagen type VIalpha 3 chain, collagen type VI alpha 5 chain, collagen type VIII alpha1 chain, and collagen type VIII alpha 2 chain; wherein the one or moreglycoproteins comprise dermatopontin, fibrillin 1, microfibril-associateprotein 4, and periostin; wherein the one or more proteoglycans compriseasporin and heparan sulfate proteoglycan 2; wherein the one or moreelastins comprise elastin isoform; wherein the one or more matrisomesecreted factors comprise chitinase, collectin subfamily member, trefoilfactor 1, and vasoactive intestinal peptide; wherein the one or more ECMregulators comprise hyaluronan binding protein 2; wherein the one ormore structural proteins comprise actin gamma 2 and myosin 11; andwherein the one or more growth factors comprise amphiregulin, basicfibroblast growth factor, bone morphogenic protein 4, bone morphogenicprotein 7, epidermal growth factor, growth differentiation factor 15,hepatocyte growth factor, insulin-like growth factor binding protein 3,and osteoprotegerin.

In some embodiments, the deconstructed matrisome comprises one or morefragments of collagens, glycoproteins, proteoglycans, elastins,matrisome secreted factors, structural proteins, growth factors, and ECMregulators, wherein the one or more collagens comprise collagen type Ialpha 1 chain, collagen type I alpha 2 chain, collagen type II alpha 1chain, collagen type III alpha 1 chain, collagen type IV alpha 1 chain,collagen type V alpha 2 chain, collagen type VI alpha 3 chain, collagentype VI alpha 5 chain; wherein the one or more glycoproteins comprisefibrillin 1, fibrillin 2, EGF-containing fibulin-like extracellularmatrix protein, laminin subunit gamma 1, prostate stem cell antigen,saposin-B-Val, and von Willebrand factor; wherein the one or moreproteoglycans comprise heparan sulfate proteoglycan; wherein the one ormore elastins comprise elastin isoform; wherein the one or morematrisome secreted factors comprise chitinase, mucin SAC, mucin 6, serumalbumin, and trefoil factor 2; wherein the one or more ECM regulatorscomprise granulin precursor; wherein the one or more structural proteinscomprise actin, keratin 1, keratin 2, keratin 9, keratin 10, myosinheavy chain 9, and tubulin beta chain; and wherein the one or moregrowth factors comprise bone morphogenic protein 4, fibroblast growthfactor 2, insulin-like growth factor binding protein 4, macrophagecolony-stimulating factor 1 receptor (CD115), and pro-epidermal growthfactor.

In some embodiments, the deconstructed matrisome comprises one or morefragments of collagens, glycoproteins, proteoglycans, elastins,matrisome secreted factors, structural proteins, growth factors, and ECMregulators, wherein the one or more collagens comprise collagen type Ialpha 1 chain, collagen type I alpha 2 chain, collagen type I alpha 3chain, collagen type II alpha 1 chain, collagen type IV alpha 1 chain,collagen type IV alpha 2 chain, collagen type IV alpha 3 chain, collagentype IV alpha 4 chain, collagen type IV alpha 5 chain, collagen type Valpha 1 chain, collagen type V alpha 2 chain, collagen type VI alpha 1chain, collagen type VI alpha 2 chain, collagen type VI alpha 3 chain,collagen type VIII alpha 1 chain, collagen type XVI alpha 1 chain, andcollagen type XXI alpha 1 chain; wherein the one or more glycoproteinscomprise fibulin 2, periostin, vitronectin, dermatopontin, lamininsubunit alpha 3, laminin subunite alpha 5, laminin subunit beta 2,laminin subunit gamma 1, and nidogen 1; wherein the one or moreproteoglycans comprise biglycan and heparan sulfate proteoglycan coreprotein; wherein the one or more elastins comprise elastin isoform;wherein the one or more matrisome secreted factors comprise hornerin;wherein the one or more ECM regulators comprise alpha 1 antitrypsin,cathepsin G, desmoplakin, junction plakoglobin, serum albumin precursor,and metalloproteinase inhibitor 3; wherein the one or more structureproteins comprise keratin 1, keratin 2, keratin 5, keratin 9, keratin10, and keratin 14; and wherein the one or more growth factors comprisebasic fibroblast growth factor, brain-derived neurotrophic factor,epidermal growth factor receptor, endocrine gland-derived vascularendothelial growth factor, growth differentiation factor 15, hepatocytegrowth factor, insulin-like growth factor binding protein 1,insulin-like growth factor binding protein 6, osteoprotegerin,platelet-derived growth factor AA, and vascular endothelial growthfactor.

The resulting mixture of matrikines in the composition may signal andmodulate unique bioactivity based on the specific mixture andconcentrations thereof. For example, the matrikines may signalproliferation, migration, protease production, apoptosis, cellinteraction, gene expression, and ECM remodeling, thereby promotingtissue repair and regeneration. In some embodiment, the deconstructedmatrisomes comprising enzymatically fragmented matrikines of the presentapplication emulate biological signaling related to tissue injury andcan trigger an immune response to promote healing. For example, thefragmentation of the matrisome may emulate a tissue injury, therebyreleasing matrikines and triggering signaling that emulates a naturalbiological response to tissue injury. Accordingly, the deconstructedmatrisome may trigger an immune response and/or promote healing byactivating signaling and cell interactions that emulate the naturalbiological response to a tissue injury. In some embodiments, where thecomposition is applied to a tissue that does not have a current orrecent injury, the deconstructed matrisome may nonetheless trigger animmune response and/or promote healing therein.

The composition described herein may be unique from TS-ECM or matrikinecompositions produced by various conventional methods by the inclusionof these various components. While conventional methods utilize intactsheets, slices, or sections, of ECM scaffold from natural tissue forcell culturing, the scaffold alone may have several drawbacks. Whileintact scaffold may be used as a covering for skin or other surfaces, itmay not penetrate and absorb through the surface due to the ECM formatas well as the fragment size.

Additionally, matrikines often display cryptic bioactivities notmanifested by the native, full-length parent proteins or molecules.While some matrikines may be active in the full-length form, manymatrikines are only active after being fragmented as described herein.In some cases, the matrikines may promote opposite activities as theparent protein. Thus, intact sheets may not present the same signalingand modulation of bioactivity as the fragmented child peptides.Exemplary active matrikines which differ in this bioactivities withrespect to their parent macromolecules are listed in Table 7(Ricard-Blum and Salza, “Matricryptins and matrikines: biologicallyactive fragments of the extracellular matrix,” Exper. Dermat., 2014, 23,457-463, incorporated by reference herein in its entirely).

Furthermore, intact sheets of scaffold may lack several components foundonly in the ECF and/or the greater matrisome. Furthermore, theconcentrations of various components in the scaffold alone may differfrom the concentrations of the same components in the whole tissue(i.e., due to the differing composition of the greater matrisome). Thematrikines described herein may be produced from TS-ECM throughprocessing ECM scaffold and tissue in a manner that does not remove orcompromise components of the extracellular environment beyond thescaffold. Therefore, the composition described herein may signal and/ormodulate bioactivity in a manner unique from an intact sheet or coveringof ECM scaffold from the same tissue source.

In contrast, the compositions described herein may compriseenzymatically fragmented deconstructed matrisome derived from ECM,thereby activating additional matrikine bioactivity and having reducedsize capable of absorbing deeper into the dermal layers of skin. Theisolated acellular extracellular matrix may be processed into fragmentsby digestion with enzymes such as proteases as would be apparent to aperson having an ordinary level of skill in the art to produce fragmentssized to be absorbed through one or more dermal layers, e.g., absorptioninto the epidermis, absorption through the epidermis and into thedermis, and/or absorption through the dermis and into the hypodermis.For example, deconstructed matrisome fragments having a size less thanabout 500 Da may be configured for absorption into the epidermis and thedermis. However, the fragments in the compositions described herein mayhave a variety of sizes, such as about 250 kDa, about 200 kDa, about 150kDa, about 100 kDa, about 50 kDa, about 25 kDa, about 10 kDa, about 5kDa, about 1 kDa, about 500 Da, about 250 Da, about 100 Da, about 50 Da,less than about Da, and/or individual values or ranges therebetween.

The matrisome may be deconstructed through one or more fragmentationprocesses as described herein, which may include chemical fragmentationsuch as enzymatic digestion (e.g., by proteases) and/or physicalfragmentation by application of a force to the source material (i.e.,biological tissue). It should be understood that the matrisome may befragmented to a greater degree than conventional processes, such asfragmentation for the purposes of producing a ECM-based cell culturesubstrate. Accordingly, by applying a variety of fragmentation processas described, the deconstructed matrisome may comprise fragmentedpeptides as small as two or more amino acids. While certain applicationssuch as cell culture only require a limited degree of fragmentation, thesubject matter disclosed herein provides for a greater degree offragmentation resulting in fragments sized and configured for absorptioninto one or more dermal layers. Furthermore, the greater degree offragmentation may emulate biological signaling related to tissue injury.For example, the fragmentation of the matrisome by the methods describedherein may emulate a tissue injury, thereby releasing matrikines andtriggering signaling that emulates a natural biological response totissue injury. Accordingly, the deconstructed matrisome may comprisefragments that are capable of triggering an immune response and/orpromoting healing by activating signaling and cell interactions thatemulate the natural biological response to a tissue injury. In someembodiments, where the composition is applied to a tissue that does nothave a current or recent injury, the deconstructed matrisome maynonetheless be capable of triggering an immune response and/or promotinghealing therein.

Furthermore, the deconstructed matrisome may have a specifiedcomposition that emulates the matrisomes found in a specific nativetissue and/or combinations thereof. As such, the composition of eachdeconstructed matrisome may vary. It should be understood that thedeconstructed matrisome includes a “complete matrisome,” i.e., a fullcomplement of naturally defined ECM components from one or morebiological tissues to provide a complete set of fragmentedmacromolecules for the purposes of promoting cell interactions andsignaling.

Each deconstructed matrisome may comprise a different combination ofproteoglycans, collagens, elastins, multiadhesive proteins, hyaluronicacid, CAMs, and additional components. Each of these components may havesubtypes, the presence of each of which may vary from one deconstructedmatrisome to another deconstructed matrisome. Each deconstructedmatrisome may be characterized by the presence or absence of one or morecomponents. Further, the concentration of each component may vary fromone deconstructed matrisome to another deconstructed matrisome. Thesevariations result in each deconstructed matrisome having unique physicalcharacteristics, such as architecture and stiffness, and unique cellinteraction characteristics, such as gene expression, ECM remodeling,and cell proliferation.

Further, the fragmented deconstructed matrisome may be decellularized,isolated and processed in a manner that does not remove or compromisecomponents of the extracellular environment beyond the scaffold.Therefore, the ECM substrates described herein include components beyondthat which is found in ECM scaffold in vivo, thereby more accuratelyemulating the in vivo extracellular environment of the tissue.

In some embodiments, the enzymatically fragmented deconstructedmatrisome may comprise additional components beyond those present in thenative extracellular matrix. In some embodiments, the enzymaticallyfragmented deconstructed matrisome may include components found in theextracellular fluid of a specific tissue. For example, a componentpresent in extracellular fluid of skin tissue may not be present in theECM scaffold thereof and may thus be added to the ECM to further emulatethe niche environment. In some embodiments, the enzymatically fragmenteddeconstructed matrisome may include one or more of amino acids, glucose,salts, vitamins, carbohydrates, proteins, peptides, trace elements,other nutrients, extracts, additives, gases, or organic compounds.Additional components for the proper growth, repair, and regeneration ofskin as would be known to one having an ordinary level of skill in theart are also contemplated herein.

As described herein, the matrikines may be configured to support tissueregeneration and healing. Further, the matrikines may be configured tofacilitate growth and proliferation of the human skin fibroblasts in amanner consistent with tissue healing. Accordingly, the matrikines mayinduce gene expression, growth factor secretion, and othercharacteristics in a manner consistent with tissue healing. However, thematrikines may be configured to support a variety of additional celltypes found in the skin, i.e., native cells.

In some embodiments, the topical formulation may have a pH of less thanabout 6.0. In some embodiments, the topical formulation has a pH of lessthan about 5.5. In some embodiments, the topical formulation has a pHselected from the group consisting of about 4.0 to about 6.0, about 4.5to about 5.0, and about 4.4 to about 4.7. The topical formulation may beconfigured to lower a pH of a skin surface to reduce or eliminatepathogen presence or growth.

The topical composition may take a variety of forms. In someembodiments, the composition is formulated in a form selected from thegroup consisting of solution, fluid, emulsion, suspension, solid,semi-solid, jelly, paste, gel, hydrogel, ointment, lotion, serum, cream,foam, mousse, liquid, spray, suspension, dispersion, powder, aerosol,film, or transdermal patches formulated as a liquid, cream, ointment,gel, aerosol, neck cream, neck lotion, body lotion, body cream, facelotion, face cream, eye lash treatment, hair moisturizer, hairconditioner, cellulite treatment, nail conditioner, gel, emulsion,silicone gel, water gel, oil-in-water emulsion, or water-in-oilemulsion. I

In some embodiments, the topical composition comprises about 0.01% byweight to about 25% by weight of deconstructed matrikines. In someembodiments, the topical composition comprises about 0.1% by weight toabout 15% by weight of deconstructed matrikines. In some embodiments,the topical composition comprises about by weight to about 2.5% byweight of deconstructed matrikines. In some embodiments, the topicalcomposition comprises deconstructed matrikines in an amount from about0.01% to about 15% by weight, about 0.01% to about 10% by weight, about0.01% to about 7.5% by weight, about 0.01% to about 5% by weight, about0.01% to about 2.5% by weight, about to about 1% by weight, about 0.01%to about 0.9% by weight, about 0.01% to about by weight, about 0.01% toabout 0.7% by weight, about 0.01% to about 0.6% by weight, about 0.01%to about 0.5% by weight, about 0.01% to about 0.4% by weight, about toabout 0.3% by weight, about 0.01% to about 0.2% by weight, about 0.01%to about by weight, about 0.01% to about 0.075% by weight, about 0.01%to about 0.05% by weight, about 0.01% to about 0.025% by weight, orindividual values or ranges therebetween.

Liquid dosage forms for topical administration may comprise diluentssuch as, for example, alcohols, glycols, oils, water, and the like. Suchcompositions may also include wetting agents or emulsifiers.

A cream can be a water-in-oil (w/o) emulsion in which an aqueous phaseis dispersed in an oil phase, or an oil-in-water (o/w) emulsion in whichan oil is dispersed within an aqueous base. An ointment generally refersto a more viscous oil-in-water cream. Traditional ointment bases (i.e.carrier) include hydrocarbons (petrolatum, beeswax, etc.) vegetableoils, fatty alcohols (cholesterol, lanolin, wool alcohol, stearylalcohol, etc.) or silicones. Insoluble solids such as starch, zincoxide, calcium carbonate, or talc can also be used in ointments andcreams. Gel forms of the compositions described above can be formed bythe entrapment of large amounts of aqueous or aqueous-alcoholic liquidsin a network of polymers or of colloidal solid particles. Such polymersor colloids (gelling or thickening agents) are typically present atconcentrations of less than 10% w/w and include carboxymethyl cellulose,hydroxypropylmethyl cellulose, hydroxyethyl cellulose, methyl cellulose,sodium alginate, alginic acid, pectin, tragacanth, carrageen, agar,clays, aluminum silicate, carbomers, and the like.

In aerosols the composition is dissolved in a propellant such asdichlorodifluoromethane, trichlorofluoromethane,dichlorotetrafluoroethane, carbon dioxide, or other suitable gas, and aco-solvent such ethanol, acetone, hexadecyl alcohol, and combinationsthereof.

Hydrogels are typically prepared by cross-linking various monomersand/or polymers to provide a three-dimensional polymer network.Non-limiting examples of polymers include, polyoxyethylene-polypropyleneblock copolymers, ionic poly saccharides, such as chitosan or sodiumalginate, cellulose, and biodegradable polymers, such as poly-lactides(PLA) and poly-glycolides (PGA), butylene succinate (PBS),polyhydroxyalkanoate (PHA), polycaprolactone acid lactone (PCL),polyhydroxybutyrate (PHB), glycolic amyl (PHV), PHB and PHV copolymer(PHBV), and poly lactic acid (PLA)-polyethylene glycol (PEG) copolymers(PLEG).

The transdermal patches can be in any conventional form such as, forexample, a strip, a gauze, a film, and the like. Patch material may benonwoven or woven (e.g., gauze dressing). Layers may also be laminatedduring processing. It may be nonocclusive or occlusive, but the latteris preferred for backing layers. The patch is preferably hermeticallysealed for storage (e.g., foil packaging). The patch can be held ontothe skin and components of the patch can be held together using variousadhesives. For example, the transdermal patch can be in the form of aBand-Aid type device, or it may be packaged in a small metal or plastic“cup”, which is strapped onto the appropriate site using an adhesive,tape, or an outer fabric or leather strap, similar to that worn as partof a watch. The entire patch may be disposable or may be refillable. Inembodiments, the formulation can be formulated with a latex polymer,wherein the formulation is applied to the skin and forms an occlusivefilm.

In some embodiments, the formulations disclosed herein can be coated onbandages, mixed with bio adhesives, or included in dressings.

In some embodiments, the formulations disclosed herein can be used incombination with a cosmetic device.

In some embodiments, the formulations disclosed herein can be used incombination with a patch.

In some embodiments, the formulation is part of an anti-aging regimen.In some embodiments, the formulation is part of regimen for after suncare. In some embodiments, the formulation is part of a photoprotectiveregimen. In some embodiments, the photoprotective regimen is a sunblockregimen or a sunscreen. In some embodiments, the formulation is part ofregimen for skin lightening. In some embodiments, the formulation ispart of regimen for skin brightening. In some embodiments, theformulation is part of regimen for acne treatment. In some embodiments,the formulation is part of regimen for inflammation treatment. In someembodiments, the formulation is part of a color cosmetic regimen. Insome embodiments, the formulation is part of a hair treatment regimen.In some embodiments, the antioxidant composition is part of a scalptreatment regimen.

In some embodiments, the topical formulation further comprises asolvent. In some embodiments, the solvent is selected from the groupconsisting of pentylene glycol, butylene glycol, water, glycols,propylene glycol, isopropylene glycol, coco-caprylate/caprate,1,2-hexanediol, and combinations thereof.

In some embodiments, the topical formulation further comprises apharmaceutical additive, a cosmetic additive, an additional agent,water, or combinations thereof. In some embodiments, the topicalformulation further comprises both a pharmaceutical additive and acosmetic additive. In some embodiments, the pharmaceutical additive, thecosmetic additive, the additional additives, or combination thereof arein a total amount of at least about 12 wt %. In some embodiments, thepharmaceutical additive, the cosmetic additive, the additional additive,or combination thereof are in a total amount selected from the groupconsisting of about 12 wt % to about 90 wt %, about 12 wt % to about 85wt %, about 12 wt % to about 80 wt %, about 15 wt % to about 90 wt %,about 15 wt % to about 85 wt %, about 15 wt % to about 80 wt %, about 15wt % to about 75 wt %, about 20 wt % to about 70 wt %, about wt % toabout 65 wt %, about 30 wt % to about 60 wt %, about 35 wt % to about 55wt %, and about 40 wt % to about 50 wt %. In some embodiments, thetopical formulation may comprise water in a percentage of at least about68 wt %. In some embodiments, the topical formulation may comprise waterin a percentage selected from the group consisting of about 68 wt % toabout 90 wt %, about 70 wt % to about 88 wt %, about 72 wt % to about 86wt %, about 74 wt % to about 84 wt %, about 76 wt % to about 82 wt %,and about 78 wt % to about wt %.

In some embodiments, the pharmaceutical additive is selected from thegroup consisting of diluents, fillers, disintegrants, binders,lubricants, surfactants, hydrophobic vehicles, water soluble vehicles,emulsifiers, buffers, humectants, moisturizers, solubilizers,preservatives, colorants, plastizers, carriers, excipients, andcombinations thereof. The person of ordinary skill in the art can referto various pharmacologic references such as, for example, ModernPharmaceutics, Banker & Rhodes, Marcel Dekker, Inc. (1979) and Goodman &Gilman's The Pharmaceutical Basis of Therapeutics, 6th Edition,MacMillan Publishing Co, New York (1980) for guidance in determining theamount of such components in the compositions and formulations ofembodiments.

In some embodiments, the cosmetic additive is selected from the groupconsisting of vitamins, cosmetic peptides, oil control agents, sensationmodifying agents, skin lightening agents, hydrating formulations,sunblock agents, a compounds that absorbs or reflects UV photons, otherskin care agents, and combinations thereof.

In some embodiments, the additional additive is selected from the groupconsisting of hydroxyacetophenone, sodium phytate, caprylic/caprictriglyceride, sodium acrylates copolymer, octyldodecanol, octyldodecylxyloside, PEG-30 dipolyhydroxystearate, Jojoba esters, Helianthus annuus(sunflower) seed wax, Acacia decurrens flower wax, polyglycerin-3,acrylamide/sodium acryloyldimethyl taurate copolymer, isohexadecane,polysorbate 80, cyclopentasiloxane, dimethicone/vinyl dimethiconecrosspolymer, ethylhexylglycerin, and combinations thereof.

In some embodiments, the topical formulation may further compriseabrasives, antiacne agents, antidandruff agents, antifungal agents,antimicrobial agents, antioxidants, toners, moisturizers, skinconditioning agents, humectants, emollients, occlusive agents, skinbleaching or lightening agents, proteins, cleaners, hair conditioners,and the like.

Abrasives may be used to remove unwanted skin such as dead skin cellsand calluses. In some embodiments, the abrasive is selected from thegroup consisting of alumina, aluminum silicate, apricot seed powder,attapulgite, avocado powder, bamboo powder, barley flour, bentonite,calcium carbonate, calcium phosphate, calcium pyrophosphate, calciumsulfate, chalk, chitin, coconut shell powder, colloidal oatmeal, comfreyleaf powder, corn cob meal or powder, corn flour, corn meal, cornstarch, diamond powder, diatomaceous earth, dicalcium phosphate,dicalcium phosphate dehydrate, egg shell powder, Fuller's earth,hydrated silica, hydroxyapatite, kaolin, kiwi seed, lauryl acrylatepolymers, loess, magnesium potassium fluorosilicate, magnesiumtrisilicate, microcrystalline cellulose, montmorillonite, Moroccan lavaclay, oat bran, oat flour, oatmeal, oyster shell powder, peach pitpowder, peanut flour, pecan shell powder, polyethylene, pumice,raspberry seed, rice bran, rye flour, sand, silica, sodium bicarbonate,sodium hydroxypropyl starch phosphate, sodium magnesium fluorosilicate,sodium silicoaluminate, soybean flour, sweet almond meal, talc, tinoxide, tricalcium phosphate, walnut shell powder, wheat bran, wheatflour, wheat powder, wheat starch, wood powder, zirconium silicate, andderivatives and combinations thereof.

Antiacne agents may be used to treat blemishes, pimples, blackheads, andwhiteheads. In some embodiments, the antiacne agent is selected from thegroup consisting of salicylic acid, benzoyl peroxide, carbamideperoxide, glycolic acid, retinal, retinol, retinaldehyde, vitamin A,vitamin A derivative, azelaic acid, or sulfur, and their derivatives andcombinations thereof.

Antidandruff agents may be used to treat dandruff, seborrheicdermatitis, or psoriasis. In some embodiments, the antidandruff agent isselected from the group consisting of coal tar, salicylic acid, seleniumsulfide, sulfur, zinc pyrithione, and their derivatives and combinationsthereof.

Antifungal agents include agents that inhibit the growth andreproduction of fungal cells or decreases the number of fungi present.In some embodiments, the antifungal agent is selected from the groupconsisting of calcium undecylenate, ketoconazol, povidone-iodine(PVP-iodine), tea tree oil, undecylenic acid, zinc undecylenate, andtheir derivatives and combinations thereof.

Antimicrobial agents include agents that kill microorganisms, prevent orinhibit microorganism growth and reproduction, or agents that helpprevent infection in minor cuts, scrapes, and burns. In someembodiments, the antimicrobial agent is selected from the groupconsisting of lower chain (C1-C₄) alcohols, quaternary ammoniumcompounds such as benzalkonium chloride and benzethonium chloride,clindamycin, methylbenzethonium chloride, hydrogen peroxide,Oligopeptide-10, phenols, tea tree oil, triclosan, povidone-iodine(PVP-Iodine), and their derivatives and combinations thereof.

Antioxidants include agents that are characterized as free radicalscavengers and help reverse skin damage caused by free radicals. In someembodiments, the antioxidant is selected from the group consisting ofacetyl cysteine, alpha lipoic acid, arbutin, ascorbic acid (vitamin C),ascorbic acid polypeptide, ascorbyl dipalmitate, ascorbyl methylsilanolpectinate, ascorbyl palmitate, ascorbyl stearate, BHA, BHT, t-butylhydroquinone, caffeic acid, Camellia sinensis oil, carotenoids, chitosanascorbate, chitosan glycolate, chitosan salicylate, chlorogenic acids,CoQ10, cortisen, cysteine, cysteine HCl, decyl mercaptomethylimidazole,diamylhydroquinone, di-t-butylhydroquinone, dicetyl thiodipropionate,dicyclopentadiene/t-butylcresol copolymer, digalloyl trioleate, dilaurylthiodipropionate, dimyristyl thiodipropionate, dioleyl tocopherylmethylsilanol, diosmine, disodium ascorbyl sulfate, disodium rutinyldisulfate, distearyl thiodipropionate, ditridecyl thiodipropionate,dodecyl gallate, dunaliella salina extract, erythorbic acid, ethylferulate, ferulic acid, hydroquinone, p-hydroxyanisole, hydroxylamineHCl, hydroxylamine sulfate, hydroxytyrosol, isooctyl thioglycolate,isoquercitrin, kojic acid, madecassicoside, magnesium ascorbate,magnesium ascorbyl phosphate, melatonin, methoxy-PEG-7 rutinylsuccinate, methylene di-t-butylcresol, methylsilanol ascorbate,nordihydroguaiaretic acid, octyl gallate, phenylthioglycloic acid,phloroglucinol, potassium ascorbyl tocopheryl phosphate, potassiumsulfite, propyl gallate, resveratrol, rosmarinic acid, rutin, sirtunis,sodium ascorbate, sodium ascorbyl/cholesteryl phosphate, sodiumbisulfite, sodium erythorbate, sodium metabisulfite, sodium sulfite,sodium thioglycolate, sorbityl furfural, tea tree oil, tetrahexyldecylascorbate, tetrahydrodiferuloylmethane, thiodiglycol, thiodiglycolamide,thiodiglycolic acid, thioglycolic acid, thiolactic acid, thiosalicylicacid, thiotaurine, tocophereth derivatives, tocopherol (vitamin E),tocophersolan, tocopheryl acetate, tocopheryl linoleate, tocopherollinoleate/oleate, tocopheryl nicotinate, tocopheryl succinate,tocoquinone, o-tolyl biguanide, tri(nonylphenyl)phosphate, ubiquinone,vitamin D, zinc dibutyldithiocarbamate, and their derivatives andcombinations thereof.

Toners include agents that create a tightening or tingling sensation onskin. In some embodiments, the toner is selected from the groupconsisting of ammonium alum, calcium chloride, calcium lactate, dimethylMEA, gallic acid, lens esculenta (lentil) seed extract, potassium alum,sodium alum, sodium aluminum chlorohydroxy lactate, sodium aluminumlactate, tannic acid, tioxolone, tranexamic acid, zinc acetate, zincchloride, zinc lactate, zinc phenolsulfonate, zinc sulfate, zirconiumchlorohydrate, witch hazel, alcohol derivatives such as denaturedalcohol and SD alcohol, aluminum derivatives such as aluminum acetate,aluminum bromohydrate, aluminum chloride, aluminum chlorohydrex,aluminum citrate, aluminum diacetate, aluminum dichlorohydrate, aluminumdichlorohydrex, aluminum glycinate, aluminum lactate, aluminumphenolsulfonate, aluminum sesquichlorohydrate, aluminumsesquichlorohydrex, and aluminum sulfate, aluminum zirconium derivativessuch as aluminum zirconium octachlorohydrex, aluminum zirconiumpentachlorohydrate, aluminum zirconium pentachlorohydrex, aluminumzirconium tetrachlorohydrate, aluminum zirconium tetrachlorohydrex,aluminum zirconium trichlorhydrate, and aluminum zirconiumtrichlorohydrex, and their derivatives and combinations thereof.

Skin conditioning agents or moisturizers can be classified intodifferent groups such as emollients, humectants, and occlusive agents.Emollients include agents that remain on the upper layers of skin andact as lubricants and improve appearance. In some embodiments, theemollient is selected from the group consisting of petrolatum,petrolatum plus volatile silicones, cold cream (USP), hydrophilicointment (USP), lanolin, glycerides, fruit oils, nut oils, vegetableoils, isopropyl palmitate, dimethicones, methicone, cyclomethicone,dormin, fatty acids, myristate derivatives like butyl myristate andmyristyl myristate, oleate derivates, C1-C4 glycols, fatty acid glycols,glycol esters, glycerine, glycerols, paraffin, rapeseed oil, long chainalcohols, olive oil, jojoba oil, castor oil, and their derivatives andcombinations thereof. Humectants include agents that increase the watercontent of the top layer of skin. In some embodiments, the humectant isselected from the group consisting of allatoin, agarose, arginine,benzyl hyaluronate, chitosan, copper, corn glycerides, gluconolactone,lactic acid, lactobionic acid, lactose, lysine, kombucha, maltitol,maltose, mannitol, propylene glycol, butylene glycol, pentylene glycol,propanediol, sodium aspartate, fructose, honey, glycerin, diglycerin,betaine, diols, hydroxyethyl urea, 1,2-hexanediol, D-ribose, glucose,sorbitol, dextrose, urea, 2-Pyrrolidone-5-Carboxylic Acid and relatedsalts, sea salt, inorganic salts of citric acid, inorganic salts oflactic acid, ectoin, glycolic acid, and their derivatives andcombinations thereof. Occlusive agents slow the evaporation of waterfrom skin. In some embodiments, the occlusive agent is selected from thegroup consisting of petrolatum, shea butter, dimethicones, plant andanimal oils such as avocado, canola, cod liver, and corn, mineral oil,olive oil, soybean oil, lanolin, glycerides, beeswax, triglycerides,long chain fatty alcohols, coco butter, coconut oil, jojoba oil,propylene glycol and their derivatives and combinations thereof.

In addition to skin conditioning agents that provide a moisturizingbenefit, there are other skin conditioning agents that improve theappearance of skin. In some embodiments, the skin conditioning agent isselected from the group consisting of cholesterol, cystine, hyaluronicacid, keratin, egg yolk, glycine, gluconolactone, lactic acid,lactobionic acid, panthenol, retinol, salicylic acid, vegetable oil,proteins, vitamins, bisabolol, ceramide, coenzyme A, lecithin and theirderivatives and combinations thereof.

Skin bleaching or lightening agents include agents that lighten pigmentin skin. The preferred skin bleaching agent is hydroquinone. In someembodiments, the brightener is selected from the group consisting ofazelaic acid, bearberry, deoxyarbuten, Glycyrrhiza glabra (Licorice)root extract, kojic acid, peat extract, and their derivatives andcombinations thereof.

Hair conditioners include agents that enhance the appearance and feel ofhair by improving a property like gloss, texture, or body. In someembodiments, the hair conditions is selected from the group consistingof lanolin, silicone, dimethicone, proteins such as amino acids,collagen, and keratin, vitamins, betaine surfactants, amine oxidesurfactants, ceramide, fatty acids, eggs, milk, natural plant and animaloils, mineral oil, olive oil, polyquaternium, and their derivatives andcombinations thereof.

Proteins include animal, plant, fungi, yeast, and bacteria proteins thathave skin health benefits. In some embodiments, the protein is selectedfrom the group consisting of collagen, keratin, soy protein, wheatprotein, bean palmitate, ascorbic acid polypeptide, the amino acids,casein, cholecalciferol polypeptide, rice protein, silk protein, glutenprotein, lysine, acetyl glucosamine, actin, actizyme, albumen,conchiorin protein, corn protein, egg protein, elastin, fibronectin,gadidae protein, hemoglobin, hexapeptide-21, lactalalbumin, lupineprotein, maple sycamore protein, milk protein, myristoyl pentapeptide-8,myristoyl tetrapeptide-8, oat protein, oligopeptide 10, palmitoylhexapeptide-14, palmitoyl oligopeptide, palmitoyl tetrapeptide-7, peaprotein, potato protein, reticulin, rice bran protein, serum protein,sweet almond protein, tetrapeptide-16, vegetable protein, yeast protein,palmitoyl oligopeptide, pantothenic acid polypeptides, milk solids,sericin, albumen, amylase, amyloglucosidase, arginine, bromelain,catalase, gelatin, zein, crystallins, cytochrome C, deoxyribonuclease,gliadin, glucose oxidase, glycoproteins, lactoferrin, lactoglubulin,lactoperoxidase, lipase, nisin, oxido reductases, papain, pepsin,subtilisin, sutilains, and their derivatives and combinations thereof.

Cleansers include agents that are used for cleaning the skin and hair bysolubilizing oil and suspending soils. Cleansers may be foaming ornon-foaming. Exemplary cleaners are typically a surfactant and can becharacterized as nonionic, anionic, or zwitterionic. In someembodiments, the cleanser is selected from the group consisting oftaurates, sulfates, sulfonates, carboxylates, sulfosuccinates,sarcosinates, zwitterionic betaines, fatty acid and fatty alcoholderivatives, and alkylpolyglucoside and amine oxide surfactants. In someembodiments, the cleansers may be combined with some abrasives such asclays and sulfurs to provide light exfoliation.

In some embodiments, the topical formulation further comprises a gellingagent. In some embodiments, the gelling agent is a water phase gellingagent. In some embodiments, the water phase gelling agent is selectedfrom the group consisting of xanthan gum, gellan gum, carrageenan,biosaccharide gum-I, sclerotium gum, pectin, pullulan, guar gum, gumarabic, chondroitin, sulfate, alginic acid, sodium hyaluronate,hydrolyzed hyaluronic acid sodium polyglutamate, chitin, chitosan,starch, and combinations thereof. In some embodiments, the gelling agentis xanthan gum.

In some embodiments, the topical formulation may further comprise an oilcontrol agent. Oil control agents are compounds useful for regulatingthe production of skin oil, or sebum, and for improving the appearanceof oily skin. In some embodiments, the oil control agent is selectedfrom the group consisting of salicylic acid, dehydroacetic acid, benzoylperoxide, vitamin B3 (for example, niacinamide), and the like, theirisomers, esters, salts and derivatives, and combinations thereof.

In some embodiments, the topical formulation may further comprise otherskin care agents selected from the group consisting of retinol,steroids, sunblock, salicylate, minocycline, antifungals, peptides,antibodies, lidocaine, and the like and combinations thereof. In someembodiments, other skin care agents include N-acyl amino acid compoundscomprising, for example, N-acyl phenylalanine, N-acyl tyrosine, and thelike, their isomers, comprising their D and L isomers, salts,derivatives, and mixtures thereof. An example of a suitable N-acyl aminoacid is N-undecylenoyl-L-phenylalanine is commercially available underthe tradename SEPIWHITE®. Other skin active agents include, but are notlimited to, Lavandox, Thallasine 2, Argireline NP, Gatuline In-Tense andGatuline Expression, Myoxinol LS 9736, Syn-ake, and Instensyl®,Sesaflash™, N-acetyl D-glucosamine, panthenol (for example, DL panthenolavailable from Alps Pharmaceutical Inc.), tocopheryl nicotinate, benzoylperoxide, 3-hydroxy benzoic acid, flavonoids (for example, flavanone,chalcone), farnesol, phytantriol, glycolic acid, lactic acid, 4-hydroxybenzoic acid, acetyl salicylic acid, 2-hydroxybutanoic acid,2-hydroxypentanoic acid, 2-hydroxyhexanoic acid, cis-retinoic acid,trans-retinoic acid, retinol, retinyl esters (for example, retinylpropionate), phytic acid, N-acetyl-L-cysteine, lipoic acid, tocopheroland its esters (for example, tocopheryl acetate: DL-a-tocopheryl acetateavailable from Eisai), azelaic acid, arachidonic acid, tetracycline,ibuprofen, naproxen, ketoprofen, hydrocortisone, acetominophen,resorcinol, phenoxyethanol, phenoxypropanol, phenoxyisopropanol,2,4,4′-trichloro-2′-hydroxy diphenyl ether, 3,4,4′-trichlorocarbanilide,octopirox, lidocaine hydrochloride, clotrimazole, miconazole,ketoconazole, neomycin sulfate, theophylline, and mixtures thereof.Further skin care agents are disclosed in US Publication No.2007/0020220A1, wherein the components/ingredients are incorporatedherein by reference in their entirety.

In some embodiments, the topical formulation may further compriseantiaging ingredients selected from the group consisting of ascorbicacid compounds, vitamin B3 compounds, azelaic acid, butylhydroxyanisole, gallic acid and its derivatives, glycyrrhizinic acid,hydroquinone, kojic acid, arbutin, mulberry extract, and combinationsthereof. In some embodiments, the topical composition or finalformulation may comprise Ovaliss((S)-5,6,6a,7-Tetrahydro-1,2,9,10-tetramethoxy-6-methyl-4H-dibenzo[de,g]quinoline,1,2-Octanediol, D-Glucopyranose, oligomeric, C10-16-alkyl glycosides,water, ethyl alcohol, and glycerin), Whey protein, MPC (Milk proteincomplex), Sesaflash (Glycerin, Acrylates copolymer, PVP/polycarbamylpolygly col ester, Hydrolyzed Sesame Protein PG-propylmethylsilanediol), Majestem (glycerin, Leontopodium Alpinum CallusCulture Extract and xanthan gum), or Idealift (butylene glycol, sorbitanlaurate, hydroxyethylcellulose, and acetyl dipeptide-1 cetyl ester).

In some embodiments, the topical formulation may further comprisessunblock agents selected from the group consisting of para-aminobenzoicacid (PABA), PABA esters (glyceryl PABA, amyldimethyl PABA andoctyldimethyl PABA), benzophenones (oxybenzone and sulisobenzone),cinnamates (octylmethoxy cinnamate and cinoxate), salicylates(homomethyl salicylate) anthranilates, TiO₂, avobenzone, bemotrizinol,bisoctrizole, 3-(4-methylbenzylidene)-camphor, cinoxate, diethylaminohydroxybenzoyl hexyl benzoate, dioxybenzone, drometrizole trisiloxane,ecamsule, ethylhexyl triazone, homosalate, menthyl anthranilate,octocrylene, octyl salicylate, iscotrizinol,isopentenyl-4-methoxycinnamate, octyl-dimethyl-p-aminobenzoic acid,octyl-methoxycinnamate, oxybenzone, polysilicone-15, trolaminesalicylate, ZnO, and combinations thereof.

In some embodiments, the topical formulation may comprise a sensationmodifying agent selected from the group of a cooling agent, a warmingagent, a relaxing or soothing agent, a stimulating or refreshing agent,and combinations thereof.

In some embodiments, the cooling agent is selected from the groupconsisting of menthol; an isomer of menthol, a menthol derivative;4-Methyl-3-(1-pyrrolidinyl)-2[5H]-furanone; WS-23, Icilin, IcilinUnilever Analog, 5-methyl-4-(1-pyrrolidinyl)-3-[2H]-furanone;4,5-dimethyl-3-(1-pyrrolidinyl)-2[5H]-furanone; isopulegol,3-(1-menthoxy)propane-1,2-diol, 3-(1-menthoxy)-2-methylpropane-1,2-diol,p-menthane-2,3-diol, p-menthane-3,8-diol,6-isopropyl-9-methyl-1,4-dioxas-piro[4,5]decane-2-methanol, menthylsuccinate and its alkaline earth metal salts, trimethylcyclohexanol,N-ethyl-2-isopropyl-5-methylcycl ohexanecarb-oxamide, Japanese mint(Mentha arvensis) oil, peppermint oil, menthone, menthone glycerolketal, menthyl lactate, 3-(1-menthoxy)ethan-1-ol,3-(1-menthoxy)propan-1-ol, 3-(1-menthoxy)butan-1-ol, 1-menthylaceticacid N-ethylamide, 1-menthyl-4-hydroxypentanoate, 1-menthyl-3-hydroxybutyrate, N,2,3-trimethyl-2-(1-methylethyl)-butanamide, spearmint oiland combination thereof.

In some embodiments, the warming agent is selected from the groupconsisting of polyhydric alcohols, capsaicin, capsicum powder, acapsicum tincture, capsicum extract, capsaicin, hamamalis,homocapsaicin, homodihydrocapsaicin, nonanoyl vanillyl amide, nonanoicacid vanillyl ether, vanillyl alcohol alkyl ether derivatives, such asvanillyl ethyl ether, vanillyl butyl ether, vanillyl pentyl ether, andvanillyl hexyl ether, isovanillyl alcohol alkyl ethers, ethylvanillylalcohol alkyl ethers, veratryl alcohol derivatives, substituted benzylalcohol derivatives, substituted benzyl alcohol alkyl ethers, vanillinpropylene glycol acetal, ethylvanillin propylene glycol acetal, gingerextract, ginger oil, gingeol, gingeron, and combination thereof.

In some embodiments, the relaxing or soothing agent is selected from thegroup consisting of herb extracts, aloe vera, alpha bisabolol,D-panthenol, allantoin, hamamelis, chamomile, yarrow; calendula,comfrey, witch hazel and other astringents, sea weed, and oat extracts;oils, selected from the group consisting of: almond oil, avocado oil,and comfrey; and essential oils, selected from the group consisting of:cardamone, eucalyptus, Mentha piperita (peppermint), hyssop, androsemary; waxy or unctuous substances selected from the group consistingof: lanolin or vaseline jelly, minerals, selected from the groupconsisting of: zinc oxide, calamine and selenium; vitamins, selectedfrom the group consisting of: tocopheryl acetate (vitamin E), andpharmaceutical agents selected from the group consisting of: analgesics,anesthetics, anti-inflammatory agents, and anti-histamines, and musclerelaxants; menthol, camphor, eugenol, eucalyptol, safrol, methylsalicylate, menthyl lactate, menthyl ethoxyacetate, menthoneglycerinacetal, 3-1-menthoxypropane-1,2-diol, ethyl 1-menthyl carbonate,(1S,3S,4R)-p-menth-8-en-3-ol, menthyl pyrrolidone carboxylate,N-substituted-p-menthane-3-carboxamides hamamelis extract, ginger oil,and combination thereof.

In some embodiments, the stimulating or refreshing agent is selectedfrom the group consisting of alcohol, L-menthol, camphor, menthe oil,capsicum extract, capsaicin, benzyl nicotinate, salicylate, glycolsalicylate, acetyl choline, serotonin, histamine, a prostaglandin, aneurotransmitter, a CNS stimulant, caffeine, quinine, and combinationthereof.

In some embodiments, the composition has a shelf life of about 1 month,about 2 months, about 3 months, about 4 months, about 5 months, about 6months, about 7 months, about 8 months, about 9 months, about 10 months,about 11 months, about 1 year, about 2 years, about 3 years, about 4years, about 5 years, about 6 years, about 7 years, about 8 years, about9 years, about 10 years, or any individual value or any range betweenany two values therein.

The topical compositions may further be configured, adapted, made and/orused in any manner described herein with respect to the method of makingand the method of using the topical compositions.

In another aspect of the present subject matter, embodiments disclosedherein are directed to methods of treating a tissue, e.g., with thetopical compositions described herein. The method may comprisingtopically administering, to a tissue of a patient, a compositioncomprising deconstructed matrisome including one or more enzymaticallyfragmented peptides (i.e., matrikines), wherein the one or moreenzymatically fragmented peptides promote one or more of tissuehomeostasis, tissue repair, and tissue regeneration. In someembodiments, the composition is topically applied to an epithelium ofthe tissue (e.g., a skin surface). In some embodiments, the compositionfurther comprises one or more excipients (e.g., a pharmaceuticallyacceptable excipient or a cosmetically acceptable excipient). In someembodiments, the deconstructed matrisome is enzymatically fragmented togenerate the one or more peptides through proteolysis (i.e., enzymaticdegradation) by proteases of one or more deconstructed matrisomecomponents. In some embodiments, the one or more enzymaticallyfragmented peptides are configured to retain cell signaling ability,thereby promoting one or more of tissue homeostasis, tissue repair, andtissue regeneration. Further, the composition may include any of thecomponents, features, characteristics, and/or variations as describedfully herein.

In some embodiments, administering the composition promotes and/orimproves at least one characteristic of the tissue.

In some embodiments, the composition increases keratin gene expression.In some embodiments, the composition increases hydration and/ormoisturization. In some embodiments, the composition increases tissueregeneration to reduce or prevent aging effects such as laxity,wrinkling, and sagging. In some embodiments, the composition increasestissue regeneration to reduce acne scarring. In some embodiments, thecomposition promotes pore size reduction and or improve skin tone.

In some embodiments, the composition reduces dermal redness. In someembodiments, the composition promotes scar healing and/or reduces scarformation. In some embodiments, the composition promotes healing and/orrecovery from a wound or burn. For example, the burn may be a burn fromheat, a chemical burn, and/or photodamage. In some embodiments, thecomposition reduces skin discoloration. For example, discoloration maybe caused by scars, redness, and/or sunspots.

In some embodiments, the composition lowers a pH of a tissue surface toreduce or eliminate pathogen presence or growth. In some embodiments,the composition may lower the pH of the tissue surface to less thanabout 6.0, less than about 5.5, less than about 5.0, less than about4.5, less than about 4.0, and/or individual values or rangestherebetween.

In embodiments described herein, improving the look or appearance ofskin is an improvement in a characteristic of the skin. In someembodiments, the characteristic of the skin is selected from the groupconsisting of firmness, elasticity, fine lines, wrinkles, skin texture,skin tone, appearance, and any combination thereof. In some embodiments,improving the look of the skin results in smoother, firmer,young-looking skin. In some embodiments, improving the look of the skinresults in a brighter complexion, improved texture, even-looking skin,and/or softer skin. In some embodiments, improving the appearance of theskin results in improvement of discoloration, disappearance ofblemishes, and/or decreased redness. In some embodiments, improving theappearance of the skin results in an anti-inflammatory effect.

In some embodiments, treating the tissue such as skin with a topicalcomposition as described herein results in an increase in expression ofkeratins, wherein the keratins comprise keratin 1, keratin 2, keratin 9,and/or keratin 10. In some embodiments, the topical composition resultsin about 300% increase in expression of keratins. However, the topicalcomposition may result in an increase in keratin expression of about50%, about 100%, about 200%, about 300%, about 400%, greater than about400%, or additional values or ranges therebetween.

In some embodiments, treating the tissue such as skin with a topicalcomposition as described herein results in an increase in cellregeneration, wherein cell regeneration is measured by restoration ofthe epithelial barrier. In some embodiments, the topical compositionresults in about 225% increase in cell regeneration. However, thetopical composition may result in an increase in cell regeneration ofabout 50%, about 100%, about 200%, about 300%, about 400%, greater thanabout 400%, or additional values or ranges therebetween.

In some embodiments, treating the tissue such as skin with a topicalcomposition as described herein results in a decrease in wound diameter.In some embodiments, the topical composition results in about 400%decrease in wound diameter. However, the topical composition may resultin a decrease in wound diameter of about 50%, about 100%, about 200%,about 300%, about 400%, about 500%, about 600%, greater than about 600%,or additional values or ranges therebetween.

In some embodiments, treating the tissue such as skin with a topicalcomposition as described herein results in accelerated healing. In someembodiments, the topical composition results in about 7 foldacceleration in healing. However, the topical composition may result inan acceleration in healing of about 2 fold, about 3 fold, about 4 fold,about 5 fold, about 6 fold, about 7 fold, about 8 fold, about 9 fold,about 10 fold, greater than about 10 fold, or additional values orranges therebetween.

In some embodiments, treating the tissue such as skin with a topicalcomposition as described herein results in a decrease in skin redness.In some embodiments, the topical composition results in about 80%decrease skin redness. However, the topical composition may result in adecrease in skin redness of about 20%, about 40%, about 60%, about 80%,about 100%, about 200%, about 300%, greater than about 300%, oradditional values or ranges therebetween.

In some embodiments, treating the tissue such as skin with a topicalcomposition as described herein results in an increase in scar healing.In some embodiments, the topical composition results in about 700%decrease skin redness. However, the topical composition may result in adecrease in skin redness of about 100%, about 200%, about 300%, about400%, about 500%, about 600%, about 700%, about 800%, about 900%, about1000% greater than about 1000%, or additional values or rangestherebetween.

In some embodiments, the composition may be configured to treat aspecific tissue condition or disease in a subject in need thereof.Accordingly, the method may comprise topically administering, to atissue of the subject, a topical composition as described herein. Insome embodiments, treating the tissue such as skin with the topicalcomposition results in an improvement in at least one symptom orcharacteristic of the tissue condition or disease.

In some embodiments, the tissue condition is selected from acne, eczema,and psoriasis. In some embodiments, the tissue condition is a fibroticskin condition selected from scleroderma, nephrogenic fibrosingdermopathy, mixed connective tissue disease, scleromyxedema,scleroderma, eosinophilic fasciitis, and combinations thereof.

While the compositions and methods herein are generally described withrespect to skin tissue, it should be understood that the composition maybe configured for application to another tissue and/or by another routeof administration. In each case, the composition may be administered inthe conventional manners accordingly.

In some embodiments the composition is a lip balm used to promote repairand regeneration of lips or other mucosal surfaces and may beadministered topically to the lips.

In some embodiments the composition is an ophthalmic solution used tominimize or reduce corneal scarring and may be administered to the eyes.

In some embodiments the composition is an edible dietary supplement usedto treat gastrointestinal health and regeneration and may beadministered orally.

In some embodiments the matrikine composition is a cosmetic product suchas a hair styling product, an anti-frizz product, an oil, a nailproduct, a beautification product, and the like.

In some embodiments the matrikine composition is a personal lubricant.

In some embodiments the composition is a personal care product such asmouth wash, sunscreen, or an after sun care product.

In some embodiments the composition is a hair care product used topromote hair or fur health and may be administered topically to the hairand/or fur. In some embodiments the composition is a hair care productused to promote growth of scalp hair, eyebrows, and eyelashes and may beadministered to the scalp, eyebrows, and/or eyelashes. In embodimentsdescribed herein, the treatment of the hair results in improvement in acharacteristic of the hair. In embodiments described herein, thecharacteristic of the hair is selected from the group consisting ofshine, texture, fullness, smoothness, density, and combinations thereof.In embodiments described herein, improving the appearance of the hairresults in smoother hair, softer hair, brighter hair, improved textureof the hair, shiner hair, fuller hair, or more vibrant hair.

In embodiments described herein, the subject is an infant, a child, anadolescent, or an adult. In embodiments described herein, the subject isan veterinary animal.

In some embodiments, the topical compositions and formulations can beapplied to the skin one, two, three, four, five or more times each day,and application can be carried out for a period of at least 1 month, 2months, 3 months, 4 months, 6 months, 8 months or 12 months.

In some embodiments, the topical compositions and formulations may beadministered once, as needed, once daily, twice daily, three times aday, once a week, twice a week, every other week, every other day, orthe like for one or more dosing cycles. A dosing cycle may compriseadministration for about 1 week, about 2 weeks, about 3 weeks, about 4weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about9 weeks, or about 10 weeks. After this cycle, a subsequent cycle maybegin approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weekslater. The treatment regime may comprise 1, 2, 3, 4, 5, or 6 cycles,each cycle being spaced apart by approximately 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 11, or 12 weeks.

In some embodiments, the methods may comprise a variety of additionalsteps including, for example, cleaning the surface tissue at the site ofapplying, abrading, microdermabrasion, toning, and the like.

The method of treating a tissue further be adapted in any mannerdescribed herein with respect to the composition and the method ofmaking the composition.

In another aspect of the present subject matter, embodiments disclosedherein are directed to methods of making the topical compositionsdescribed herein. A wide variety of methods may be used for preparingthe compositions and formulations described herein. Broadly speaking,the compositions may be prepared by combining the components of theformulation, as described herein, at a temperature and for a timesufficient to provide a pharmaceutically or cosmetically acceptablecomposition.

FIG. 1 depicts a diagram of an illustrative method of making the topicalcomposition in accordance with an embodiment. The method may comprisesproviding 105 a tissue, isolating 110 a decellularized, acellulartissue-specific extracellular matrix from biological tissue, fragmentingand solubilizing 115 the tissue-specific extracellular matrix using oneor more enzymes to produce a deconstructed matrisome solution includingone or more enzymatically fragmented peptides (i.e., matrikines), andcombining 120 the matrix solution with one or more excipients (e.g., apharmaceutically acceptable excipient or a cosmetically acceptableexcipient) to form the topical composition. In some embodiments, the oneor more peptides promote one or more of tissue homeostasis, tissuerepair, and tissue regeneration. In some embodiments, the composition isconfigured for topical application to an epithelium of the tissue (e.g.,a skin surface). In some embodiments, the deconstructed matrisome isfragmented to generate the one or more peptides through proteolysis(i.e., enzymatic degradation) of one or more deconstructed matrisomecomponents. Further, the composition may include any of the components,features, characteristics, and/or variations as described fully herein.

According to an exemplary embodiment, tissue is procured and immediatelyfrozen and prepared for sectioning. Frozen blocks are then sectionedlongitudinally into thin (200 μm-1 mm) slices showing the entirecross-section of the tissue. Portions of the tissue may be dissected andseparated from the thin slices prior to decellularization. The tissuesare treated using a sequence of chemical, detergent, and enzymaticwashes. Each wash is followed by de-ionized water washes. In someembodiments, each region is decellularized by serial washes up to 12hours followed by enzymatic digestions.

The described process may be modified or adapted for various tissuesdescribed herein. Tissue sections are decellularized by the introductionof one or more of deionized water, hypertonic salines, enzymes,detergents, and acids. In an exemplary embodiment, tissue sections aredecellularized by using a sequence of chemical, detergent, and enzymaticwashes. Each wash may be followed by de-ionized water washes.

Following decellularization, the decellularized material is snap frozenin liquid nitrogen, pulverized, milled, and lyophilized to obtain a fineECM powder. In some embodiments, the ECM powder is digested using anenzymatic agent. In some embodiments an ECM solution is produced fromthe ECM powder. The resulting digest may neutralized, frozen, and thawedto obtain ECM solution. In some embodiments, the solubilizing step maynot be performed and the ECM material may be utilized in its powderform.

The ECM solution or ECM powder may be combined with an excipient such asa pharmaceutically acceptable excipient and/or a cosmetically acceptableexcipient to produce the topical composition. The ECM material and theexcipient may be combined, mixed, and/or homogenized. Further, anynumber of additional materials, such as the agents and/or inactiveingredients described herein with respect to the topical formulation.

In some embodiments, the ECM solution may be reconstituted into anotherform before being combined with additional materials. The ECM materialmay be reconstituted into a hydrogel or another format by adding areagent such as a buffer to adjust the ionic strength and the pH of thesolution. In some embodiments, the reagent comprises one or more of aneutral buffer, a basic buffer, a base, and an acid. For example, aneutral buffer may comprise Phosphate Buffered Saline (PBS), TAPSO(3-[N-tris(hydroxymethyl)methylamino]-2-hydroxypropanesulfonic acid),HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfoni c acid), TES(2-[[1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl]amino]ethanesulfonicacid), and/or MOPS (3-(N-morpholino)propanesulfonic acid). For example,a basic buffer may comprise carbonate bicarbonate, TAPS([tris(hydroxymethyl)methylamino]propanesulfonic acid), Bicine(2-(bis(2-hydroxyethyl)amino)acetic acid), Tris(tris(hydroxymethyl)aminomethane), and/or Tricine(N-[tris(hydroxymethyl))methyl]glycine). For example, a base maycomprise Sodium Hydroxide (NaOH). For example, an acid may compriseHydrochloric Acid (HCl) or Acetic Acid. In additional embodiments, thereagent may comprise deionized water. However, additional or alternativereagents may be provided to convert the ECM material into various forms,as would be known to a person having an ordinary level of skill in theart. In still additional embodiments, a reagent is not required.

In some embodiments, the process may be further adapted based on theproperties of the tissue. In some embodiments, a higher content ofconnective tissue and/or the greater mechanical stiffness may requirelonger digestion than would be required for other tissues. In someembodiments, to form a solution as described, the ECM powder is digestedwith an enzymatic agent for about 1 hour, about 2 hours, about 3 hours,about 4 hours, about 5 hours, greater than about 5 hours, or individualvalues or ranges therebetween.

In some embodiments, a plurality of deconstructed matrisome solutionsmay be formed by the manner described herein and used to form thetopical composition. In some embodiments, the topical composition maycomprise one, two, three, four, five, or more different deconstructedmatrisome solutions.

In some embodiments, the topical composition has a shelf life of about 1month, about 2 months, about 3 months, about 4 months, about 5 months,about 6 months, about 7 months, about 8 months, about 9 months, about 10months, about 11 months, about 1 year, about 2 years, about 3 years,about 4 years, about 5 years, about 6 years, about 7 years, about 8years, about 9 years, about 10 years, greater than about 10 years, orany individual value or any range between any two values therein.

The topical composition may be characterized by any of the components,concentrations thereof, and/or changes thereof from normal as summarizedin Table 1, Table 2, and Table 3, Table 4, Table 5 and Table 6. However,these compositions are exemplary in nature and the ECM profiles may varytherefrom as to any number of components. For example, the compositionof the substrate may vary from the described concentration values and/orranges by about 10%, about 20%, about 30%, greater than 30%, orindividual values or ranges therebetween.

As described herein, the matrikines in the topical composition may beconfigured to support tissue regeneration and healing. Further, thematrikines may be configured to facilitate growth and proliferation ofthe human skin fibroblasts in a manner consistent with tissue healing.Accordingly, the matrikines may induce gene expression, growth factorsecretion, and other characteristics in a manner consistent with tissuehealing. However, the matrikines may be configured to support a varietyof additional cell types found in the skin, i.e., native cells.

The method of making a ECM substrate may further be adapted in anymanner described herein with respect to the ECM substrate and the methodof using the ECM substrate.

The subject matter is now described with reference to the followingexamples. These examples are provided for the purpose of illustrationonly and the claims should in no way be construed as being limited tothese examples, but rather should be construed to encompass any and allvariations which become evident as a result of the teaching providedherein. Those of skill in the art will readily recognize a variety ofnon-critical parameters that could be changed or modified to yieldessentially similar results.

Examples

The disclosures of each and every patent, patent application,publication, and accession number cited herein are hereby incorporatedherein by reference in their entirety.

While present disclosure has been disclosed with reference to variousembodiments, it is apparent that other embodiments and variations ofthese may be devised by others skilled in the art without departing fromthe true spirit and scope of the disclosure. The appended claims areintended to be construed to include all such embodiments and equivalentvariations.

Example 1: Gene Expression of Normal Human Dermal Fibroblast (NHDF)Cultured in Different Deconstructed Matrisome Components

NHDFs were cultured on plastic as well as in blood vessel-, skin-,liver-, intestine-, and cartilage-specific extracellular matrixcomponents, as seen by light microscope (FIG. 2 ) and immunofluorescentmicroscopy (FIG. 3 ; SDS-PAGE of blood vessel, BV, and skin, SK,extracellular matrix shown in FIG. 4 ). Following RNA extraction andqPCR, the presence of genes related to wound-healing and scarring(including EGFR, IL1, TGFb1, COLA1, PDGFC, PDGFRB, FGF2, MMP2) werequantified. Data from blood vessel and skin were averaged across threelots for both of these tissue types. NHDFs showed decreased expressionof wound healing genes across all tissue-specific component conditionsin comparison to plastic (no deconstructed matrisome components) (FIG. 5). Interleukin-1 gene expression was reduced in the presence ofdeconstructed matrisome components suggesting that these components mayinhibit inflammatory responses. Genes may be induced or expresseddifferently in each specific tissue type.

Example 2: Characterization of Matrikine Formula 1

Biochemical analysis of blood vessel and skin extracellular matrixcomponents, or matrikines. Extracellular matrix components from bloodvessel and skin were analyzed for protein expression. Minimalvariability across lots within each tissue type was observed (FIG. 6 ).

Example 3: Matrikynes Increase Skin Healing

Interactions between the matrikines and the microbiome are instrumentalto healthy barrier function and protection from pathogens. The intrinsicbioactivity of matrikines regulate immunologic response, instruct dermalepithelial cell organization and restore normal skin tissuearchitecture. Matrikynes® lower skin surface pH (5.5) to make skininhospitable to pathogens and produce antimicrobial peptides that limitgrowth of pathogenic microbes on skin. Matrikynes® containingextracellular matrix fragments smaller than 500 Da allow for penetrationinto the dermis (FIG. 7 ). The experiments and measurements herequantify this regeneration.

Matrikynes®, according to Formula 1 (Table 1), promote epithelialregeneration, which is measured by an increase in keratin expression(FIG. 7 ). Matrikynes® improve skin repair and healing and reduce theappearance of scars. Application of Matrikynes® showed about an 86%reduction in redness as measured by mean pixels above threshold overthirty six weeks (FIG. 8 ). Matrikynes® reduce the size of wounds ascompared to untreated tissue by 421% with 7.3 times faster healing (FIG.9 ). In summary, treatment with Matrikynes® leads to a 316% increase inkeratin expression, a 227% increase in cellular regeneration compared tountreated tissue, a 421% increase in wound closure, an 86% decrease inwound redness, and a 726% increase in skin healing (FIG. 10 ).

Example 4: Method of Making Matrikynes®

Tissue-specific decellularized extracellular matrix can be formulatedinto skin care formulations for topical application. The first step increating any of these products is to isolate tissue and collect tissuesamples. The tissue from these samples then undergoes a cell removalprocess, or decellularization, and the extracellular matrix is themisolated away from the rest of the cellular components. Once theextracellular matrix is isolated it is then fragmented by enzymaticdegradation using a number of proteases and processed into a powder forreconstitution in a topical formulation for skin treatment.

Example 5: Matrikynes Modulate Cellular Activity

Primary human epithelial cells were cultured for 4 days (FIG. 11 ).Scratch assay was performed with Matrikynes® (Formula 1, Table 1), andwithout Matrikynes® (control) to evaluate effects of Matrikynes®, whichafter 24 hours increased cellular migration, proliferation, and woundclosure compared to untreated controls (scale bar: 100 μm).

Example 6: Human Repeat Insult Patch Test (HRIPT)

Substances that come in contact with human skin must be evaluated forpropensity to irritate and/or sensitize. A reproducible, standardized,quantitative patch evaluation procedure must be used to demonstrate thata particular material can be applied safely to human skin withoutsignificant risk of adverse reactions.

Sensitization was determined by repeated topical applications to theskin of human subjects under controlled patch test conditions. Repeatedinsult patch evaluation is a predictive patch study that can detectirritant reactions and weak sensitizers that require multipleapplications to induce a cell-mediated (type IV) immune responsesufficient to cause an allergic reaction. A reproducible, standardized,quantitative patch evaluation procedure was used to demonstrate thatMatrikynes® at 2.5% by weight can be applied safely to human skinwithout adverse reaction (FIGS. 12A and 12B; N=106 human subjects).Matrikynes® of Formula I (Table 1) were used in the HRIPT.

Example 7: Case Studies in Wound and Scar Healing

Superficial Burn: Methods: Base cream containing Matrikynes® (0.1% byweight; Formula 1, Table 1) was applied twice daily for four (4) weeksto the site of a superficial burn on the index finger. No othermedications or products were used. Results: Gross photography showedreduced redness, skin repair, and scar-free healing after four (4) weeks(FIG. 13 ).

Wound Healing: Methods: Base cream containing Matrikynes® (0.1% byweight; Formula 1, Table 1) was applied twice daily for eight (8) weeksto the site of a wound closed with surgical suture on the palm of thehand. No other medications or products were used. Results: Grossphotography showed reduced redness and healing after eight (8) weeks(FIG. 14 ).

Wound Healing with Surgical Suture: Methods: Base cream containingMatrikynes® (0.1% by weight; Formula 1, Table 1) was applied twice dailyover the course of five (5) weeks to the site of a wound closed withsurgical suture on the lower leg. No other medications or products wereused. Results: Gross photography showed reduced redness and healingwithout scar after five (5) weeks (FIG. 15 ).

Scar Reduction: Methods: Base cream containing Matrikynes® (0.1% byweight; Formula 1, Table 1) was applied twice daily for twenty (20)weeks to the site of post-surgical fine line scars on the upper arm. Noother medications or products were used. Results: Gross photographyshowed reduced raise of scars, smoother skin texture at the scar sites,and improved appearance after twenty (20) weeks (FIG. 16 ).

Scar Reduction: Methods: Base cream containing Matrikynes® (0.1% byweight; Formula 1, Table 1) was applied twice daily for five (5) weeksto a Caesarian section scar. No other medications or products were used.Results: Gross photography showed reduced raise and length of scar,reduced discoloration, and improved appearance after five (5) weeks(FIG. 17 ).

Scar Reduction: Methods: Base cream containing Matrikynes® (0.1% byweight; Formula 1, Table 1) was applied twice daily for eight (8) weeksto a seven-year-old ACL knee surgery scar. No other medications orproducts were used. Results: Gross photography showed reduced raise andlength of scar, reduced wrinkles, and improved appearance after eight(8) weeks (FIG. 18 ).

Scar Reduction: Methods: Base cream containing Matrikynes® (0.1% byweight; Formula 1, Table 1) was applied twice daily for twelve (12)weeks to a two-year-old hip replacement surgery scar in a 61 year oldfemale subject. No other medications or products were used. Results:Gross photography showed visible diminished appearance of scar aftertwelve (12) weeks (FIG. 19 ).

Example 8: Case Studies in Treating Skin Conditions

Acne vulgaris study 1: Methods: Base cream containing Matrikynes® (0.1%by weight; Formula 1, Table 1) was applied twice daily for six (6) weeksto areas of the face with acne lesions. No other medications or productswere used. Results: Gross photography showed resolution of acne lesions,reduced post-inflammatory erythema, and improved skin tone after six (6)weeks (FIG. 20 ).

Acne vulgaris study 2: Methods: Base cream containing Matrikynes® (0.1%by weight; Formula 1, Table 1) was applied twice daily for two (2) weeksto areas of the face with acne lesions. No other medications or productswere used. Results: Gross photography showed resolution of acne lesions,reduced post-inflammatory erythema, and improved skin tone and textureafter two (2) weeks (FIG. 21 ).

Anti-aging and anti-wrinkle: Methods: Base cream containing Matrikynes®(0.1% by weight; Formula 1, Table 1) was applied twice daily for six (6)weeks to the face. No other medications or products were used. Results:Gross photography showed reduced appearance of: photodamage, redness,and fine lines after two (2) weeks (FIG. 22 ).

Subjects using Matrikynes® (0.1% by weight; Formula 1, Table 1) fortreatment of skin conditions reported high levels of satisfaction (FIG.23 ). Subjects found that skin felt softer, skin felt smoothed, skintexture felt improved, and skin looked healthier. Subjects agreed thatthe product reduced appearance of redness or discoloration and improvedappearance of blemishes and scars (FIG. 23 ).

Example 9: Anti Aging Properties of Matrikynes®

Objective and Methods: Subjects aged 35 to 65 years of age, will beobserved for skin changes after treatment with the Matrikyne® product.The clinical study will evaluate the parameters listed below. A baselinefor each of the parameters measured will be established after a 5 daywashout period. Subjects will then be given the Matrikyne® product touse according to instructions for the duration of the study. Subjectswill return to the clinic after 1 hour, 4 hours, 8 hours, 4 weeks and 8weeks for evaluation.

Parameters: 1) reduction in the appearance of fine lines and wrinkles(globally), measured by VAESTRO analysis of photos obtained withCanfield VISIA CR; 2) subjective, measured by a questionnaire; 3)improvement in skin firmness and elasticity, measured by cutometer; 4)improvement in skin hydration/moisturization, measured by corneometer;5) improvement in skin barrier function, measured by TEWL; 6)improvement in the appearance of hyperpigmentation/age spots, measuredby VAESTRO analysis of photos with Canfield VISIA CR; 7) improvement inskin density, measured by ultrasound; 8) reduction in the appearance offine lines and wrinkles in the Crow's feet, measured by primos 3D; 9)repair of skin barrier function, measured by TWEL with tape stripping.

Anticipated Results: Subjects treated with Matrikyne® will demonstrate areduction in the appearance of fine lines and wrinkles (globally as wellas Crow's feet), will subjectively report an improvement in skincondition, an improvement in skin firmness and elasticity, animprovement in skin hydration/moisturization, an improvement in skinbarrier function, an improvement in the appearance ofhyperpigmentation/age spots, an improvement in skin density, and repairof skin barrier function.

What is claimed is:
 1. A composition for topical administration to anepithelium, the composition comprising: a deconstructed matrisomeincluding one or more enzymatically fragmented peptides derived from atleast one biological tissue; and one or more pharmaceutically acceptableor cosmetically acceptable excipients, wherein the deconstructedmatrisome is in an amount from about 0.1% by weight to about 15% byweight of the composition; and wherein the one or more enzymaticallyfragmented peptides are configured to retain cell signaling ability,thereby promoting one or more of tissue homeostasis, tissue repair, andtissue regeneration.
 2. The composition of claim 1, wherein thedeconstructed matrisome is in an amount from about 0.1% by weight toabout 2.5% by weight of the composition.
 3. The composition of claim 1,wherein the enzymatically fragmented peptides are sized and configuredto be absorbed through one or more dermal layers.
 4. The composition ofclaim 3, wherein the one or more dermal layers comprise epidermis anddermis.
 5. The composition of claim 1, where in the enzymaticallyfragmented peptides have a size of less than about 500 Da.
 6. Thecomposition of claim 6, where in the enzymatically fragmented peptideshave a size of less than about 250 Da.
 7. The composition of claim 1,wherein the composition has a pH of less than about 6.0.
 8. Thecomposition of claim 1, wherein the deconstructed matrisome comprisesone or more of a solution and a powder.
 9. The composition of claim 1,wherein the deconstructed matrisome comprises one or more fragments ofcollagens, glycoproteins, proteoglycans, glycosaminoglycans, laminins,extracellular matrix associated proteins, soluble growth factors,inflammatory cytokines and chemokines, and immune mediators.
 10. Thecomposition of claim 9, wherein the fragments of collagens are in anamount from about 400 ng/mL to about 9700 ng/mL.
 11. The composition ofclaim 9, wherein the fragments of collagens comprise collagen type IV inan amount from about 2 ng/mL to about 24 ng/mL.
 12. The composition ofclaim 9, wherein the fragments of glycosaminoglycans are in an amountfrom about 3 μg/mL to about 170 ng/mL.
 13. The composition of claim 1,wherein the deconstructed matrisome comprises one or more fragments ofcollagens, glycoproteins, proteoglycans, elastins, matrisome secretedfactors, structural proteins, growth factors, and ECM regulators. 14.The composition of claim 13, wherein the fragments of collagens are inan amount from about 400 ng/mL to about 9700 ng/mL.
 15. The compositionof claim 13, wherein the fragments of elastins are in an amount of about40 ng/mL to about 3000 ng/mL.
 16. The composition of claim 13, whereinthe one or more fragments of collagens comprise collagen type I alpha 1chain, collagen type III alpha 1 chain, and collagen type V alpha 2chain; wherein one or more fragments of glycoproteins comprise fibrillarcollagen NC1 domain-containing protein, fibrillin 1, and microfibrilassociated protein 4; wherein the one or more fragments of proteoglycanscomprise heparan sulfate proteoglycan 2; wherein the one or morefragments of elastins comprise elastin isoform; wherein the one or morefragments of structural proteins comprise actin gamma 2 and filamin A;and wherein the one or more fragments of growth factors comprise latenttransforming growth factor beta binding protein
 4. 17. The compositionof claim 13, wherein the one or more fragments of collagens comprisecollagen type I alpha 1 chain, collagen type I alpha 2 chain, collagentype II alpha 1 chain, collagen type III alpha 1 chain, collagen type Valpha 1 chain, collagen type V alpha 2 chain, collagen type VI alpha 2chain, collagen type VI alpha 3 chain, collagen type VIII alpha 1 chain,collagen type IX alpha 2 chain, collagen type XI alpha 1 chain, collagentype XI alpha 2 chain, collagen type XII alpha 2 chain, and collagentype XIV alpha 1 chain; wherein the one or more fragments ofglycoproteins comprise fibrillin 1, adipocyte enhancer binding protein1, alpha-2-Heremans-Schmid glycoprotein, biglycan, extracellular matrixprotein 2, fibrinogen beta chain, fibrinogen gamma chain, fibronectin 1,osteonectin, periostin, tenascin C, tenascin N, thrombospondin 1,transforming growth factor beta induced, and vitronectin; wherein theone or more fragments of proteoglycans comprise heparan sulfateproteoglycan 2, aggrecan core protein, asporin, decorin, fibromodulin,lumican, mimecan, osteoglycan, osteomodulin, and proline/arginine-richend leucine-rich repeat protein; wherein the one or more fragments ofelastins comprise elastin; wherein the one or more fragments ofmatrisome secreted factors comprise albumin, annexin A2, chitinase,collectin subfamily member 12, creatine kinase B, olfactomedin; whereinthe one or more fragments of ECM regulators are coagulation factor IX,coagulation factor X, inter-alpha (globulin) inhibitor H4, prothrombin,and serpin peptidase inhibitor Glade F; and wherein the one or morefragments of structural proteins are actin gamma 2 and vimentin.
 18. Thecomposition of claim 13, wherein the one or more fragments of collagenscomprise collagen type I alpha 1 chain, collagen type I alpha 2 chain,collagen type II alpha 1 chain, collagen type III alpha 1 chain,collagen type IV alpha 1 chain, collagen type IV alpha 2 chain, collagentype V alpha 1 chain, collagen type V alpha 2 chain, collagen type VIalpha 2 chain, collagen type VI alpha 3 chain, collagen type VI alpha 5chain, collagen type VIII alpha 1 chain, and collagen type VIII alpha 2chain; wherein the one or more fragments of glycoproteins comprisedermatopontin, fibrillin 1, microfibril-associate protein 4, andperiostin; wherein the one or more fragments of proteoglycans compriseasporin and heparan sulfate proteoglycan 2; wherein the one or morefragments of elastins comprise elastin isoform; wherein the one or morefragments of matrisome secreted factors comprise chitinase, collectinsubfamily member, trefoil factor 1, and vasoactive intestinal peptide;wherein the one or more fragments of ECM regulators comprise hyaluronanbinding protein 2; wherein the one or more fragments of structuralproteins comprise actin gamma 2 and myosin 11; and wherein the one ormore fragments of growth factors comprise amphiregulin, basic fibroblastgrowth factor, bone morphogenic protein 4, bone morphogenic protein 7,epidermal growth factor, growth differentiation factor 15, hepatocytegrowth factor, insulin-like growth factor binding protein 3, andosteoprotegerin.
 19. The composition of claim 13, wherein the one ormore fragments of collagens comprise collagen type I alpha 1 chain,collagen type I alpha 2 chain, collagen type II alpha 1 chain, collagentype III alpha 1 chain, collagen type IV alpha 1 chain, collagen type Valpha 2 chain, collagen type VI alpha 3 chain, collagen type VI alpha 5chain; wherein the one or more fragments of glycoproteins comprisefibrillin 1, fibrillin 2, EGF-containing fibulin-like extracellularmatrix protein, laminin subunit gamma 1, prostate stem cell antigen,saposin-B-Val, and von Willebrand factor; wherein the one or morefragments of proteoglycans comprise heparan sulfate proteoglycan;wherein the one or more fragments of elastins comprise elastin isoform;wherein the one or more fragments of matrisome secreted factors comprisechitinase, mucin SAC, mucin 6, serum albumin, and trefoil factor 2;wherein the one or more fragments of ECM regulators comprise granulinprecursor; wherein the one or more fragments of structural proteinscomprise actin, keratin 1, keratin 2, keratin 9, keratin 10, myosinheavy chain 9, and tubulin beta chain; and wherein the one or morefragments of growth factors comprise bone morphogenic protein 4,fibroblast growth factor 2, insulin-like growth factor binding protein4, macrophage colony-stimulating factor 1 receptor (CD115), andpro-epidermal growth factor.
 20. The composition of claim 13, whereinthe one or more fragments of collagens comprise collagen type I alpha 1chain, collagen type I alpha 2 chain, collagen type I alpha 3 chain,collagen type II alpha 1 chain, collagen type IV alpha 1 chain, collagentype IV alpha 2 chain, collagen type IV alpha 3 chain, collagen type IValpha 4 chain, collagen type IV alpha 5 chain, collagen type V alpha 1chain, collagen type V alpha 2 chain, collagen type VI alpha 1 chain,collagen type VI alpha 2 chain, collagen type VI alpha 3 chain, collagentype VIII alpha 1 chain, collagen type XVI alpha 1 chain, and collagentype XXI alpha 1 chain; wherein the one or more fragments ofglycoproteins comprise fibulin 2, periostin, vitronectin, dermatopontin,laminin subunit alpha 3, laminin subunit alpha 5, laminin subunit beta2, laminin subunit gamma 1, and nidogen 1; wherein the one or morefragments of proteoglycans comprise biglycan and heparan sulfateproteoglycan core protein; wherein the one or more fragments of elastinscomprise elastin isoform; wherein the one or more fragments of matrisomesecreted factors comprise hornerin; wherein the one or more fragments ofECM regulators comprise alpha 1 antitrypsin, cathepsin G, desmoplakin,junction plakoglobin, serum albumin precursor, and metalloproteinaseinhibitor 3; wherein the one or more fragments of structure proteinscomprise keratin 1, keratin 2, keratin 5, keratin 9, keratin 10, andkeratin 14; and wherein the one or more fragments of growth factorscomprise basic fibroblast growth factor, brain-derived neurotrophicfactor, epidermal growth factor receptor, endocrine gland-derivedvascular endothelial growth factor, growth differentiation factor 15,hepatocyte growth factor, insulin-like growth factor binding protein 1,insulin-like growth factor binding protein 6, osteoprotegerin,platelet-derived growth factor AA, and vascular endothelial growthfactor.
 21. A method of promoting homeostasis, repair, or regenerationin a target tissue, the method comprising: topically administering acomposition to an epithelium of the target tissue, the compositioncomprising: a deconstructed matrisome including one or moreenzymatically fragmented peptides derived from at least one biologicaltissue, and one or more pharmaceutically acceptable or cosmeticallyacceptable excipients, wherein the deconstructed matrisome is in anamount from about 0.1% by weight to about 15% by weight of thecomposition; and wherein the one or more enzymatically fragmentedpeptides are configured to retain cell signaling ability, therebypromoting one or more of tissue homeostasis, tissue repair, and tissueregeneration in the target tissue.
 22. The method of claim 21, whereinthe tissue exhibits one or more of acute injury, wound, scarring, acne,eczema, and psoriasis.
 23. The method of claim 21, wherein topicallyadministering the composition results in an increase in expression ofkeratins.
 24. The method of claim 21, wherein the keratins comprise oneor more of keratin 1, keratin 2, keratin 9, and keratin
 10. 25. Themethod of claim 21, wherein topically administering the compositionresults in an increase in cell regeneration.
 26. The method of claim 21,wherein the tissue is a skin tissue, and wherein topically administeringthe composition results in a decrease in skin redness.
 27. The method ofclaim 21, wherein the tissue comprises a wound, and wherein topicallyadministering the composition results in a decrease in a diameter of thewound.
 28. The method of claim 21, wherein the tissue comprises aninjury, and wherein topically administering the composition results in areduction in the formation and appearance of scars.
 29. The method ofclaim 21, wherein the tissue comprises scarring, and wherein topicallyadministering the composition results in an increase in scar healing.30. A method of increasing keratin gene expression in a target tissue,the method comprising: topically administering a composition to anepithelium of the target tissue, the composition comprising: adeconstructed matrisome including one or more enzymatically fragmentedpeptides derived from at least one biological tissue, and one or morepharmaceutically acceptable or cosmetically acceptable excipients,wherein the deconstructed matrisome is in an amount of about 0.1% byweight to about 15% by weight of the composition; and wherein the one ormore enzymatically fragmented peptides are configured to retain cellsignaling ability, thereby reducing or preventing one or more of laxity,wrinkling, and sagging in the target tissue.
 31. A method of reducingdermal redness in a target tissue, the method comprising: topicallyadministering a composition to an epithelium of the target tissue, thecomposition comprising: a deconstructed matrisome including one or moreenzymatically fragmented peptides derived from at least one biologicaltissue, and one or more pharmaceutically acceptable or cosmeticallyacceptable excipients, wherein the deconstructed matrisome is in anamount of about 0.1% by weight to about 15% by weight of thecomposition; and wherein the one or more enzymatically fragmentedpeptides are configured to retain cell signaling ability, therebypromoting healing or recovery in the target tissue from one or more ofscar formation, a wound, and a burn.
 32. A method of lowering the pH ofa surface of a target tissue, the method comprising: topicallyadministering a composition to an epithelium of the target tissue, thecomposition comprising: a deconstructed matrisome including one or moreenzymatically fragmented peptides derived from at least one biologicaltissue, and one or more pharmaceutically acceptable or cosmeticallyacceptable excipients, wherein the deconstructed matrisome is in anamount of about 0.1% by weight to about 15% by weight of thecomposition; and wherein the one or more enzymatically fragmentedpeptides are configured to retain cell signaling ability, therebylowering the pH of the surface and reducing presence or growth of apathogen on the surface of the target tissue.
 33. A method of improvinga characteristic of a target skin tissue, the method comprising:topically administering a composition to an epithelium of the targetskin tissue, the composition comprising: a deconstructed matrisomeincluding one or more enzymatically fragmented peptides derived from atleast one biological tissue, and one or more pharmaceutically acceptableor cosmetically acceptable excipients, wherein the deconstructedmatrisome is in an amount of about 0.1% by weight to about 15% by weightof the composition; wherein the one or more enzymatically fragmentedpeptides are configured to retain cell signaling ability, therebyimproving the characteristic of the target skin tissue; and wherein thecharacteristic of the target skin tissue is selected from the groupconsisting of firmness, elasticity, fine lines, wrinkles, skin texture,skin tone, and appearance.
 34. A method of increasing cell regenerationin a target tissue, the method comprising: topically administering acomposition to an epithelium of the tissue, the composition comprising:a deconstructed matrisome including one or more enzymatically fragmentedpeptides derived from at least one biological tissue, and one or morepharmaceutically acceptable or cosmetically acceptable excipients,wherein the deconstructed matrisome is in an amount of about 0.1% byweight to about 15% by weight of the composition; and wherein the one ormore enzymatically fragmented peptides are configured to retain cellsignaling ability, thereby restoring an epithelial barrier of the targettissue.